Chemotaxis to cyclic AMP and folic acid is mediated by different G proteins in Dictyostelium discoideum

NARCIS (Netherlands)

Kesbeke, Fanja; Haastert, Peter J.M. van; Wit, René J.W. de; Snaar-Jagalska, B. Ewa

1990-01-01

Mutant Frigid A (fgdA) of Dictyostelium discoideum is defective in a functional Gα2 subunit of a G protein and is characterized by a complete blockade of the cyclic AMP-mediated sensory transduction steps, including cyclic AMP relay, chemotaxis and the cyclic GMP response. Folic acid-mediated

Transient Kinetics of a cGMP-dependent cGMP-specific Phosphodiesterase from Dictyostelium discoideum

NARCIS (Netherlands)

Haastert, Peter J.M. van; Lookeren Campagne, Michiel M. van

1984-01-01

Chemotactic stimulation of Dictyostelium discoideum cells induces a fast transient increase of cGMP levels which reach a peak at 10 s. Prestimulation levels are recovered in ~30 s, which is achieved mainly by the action of a guanosine 3',5'-monophosphate cGMP-specific phosphodiesterase. This enzyme

RNAi silenced Dd-grp94 (Dictyostelium discoideum glucose-regulated protein 94 kDa) cell lines in Dictyostelium exhibit marked reduction in growth rate and delay in development.

Science.gov (United States)

Baviskar, Sandhya N; Shields, Malcolm S

2010-01-01

Glucose-regulated 94 kDa protein (Grp94) is a resident of the endoplasmic reticulum (ER) of multicellular eukaryotes. It is a constitutively expressed protein that is overexpressed in certain abnormal conditions of the cell such as depletion of glucose and calcium, and low oxygen and pH. The protein is also implicated in diseased conditions like cancer and Alzheimer's disease. In this study, the consequences of downregulation of Grp94 were investigated at both unicellular and multicellular stages of Dictyostelium discoideum. Previous studies have shown the expression of Dd-Grp94 (Dictyostelium discoideum glucose-regulated 94 kDa protein) in wild-type cells varies during development, and overexpression of Dd-Grp94 leads to abnormal cell shape and inhibition of development (i.e., formation of fruiting bodies). Grp94 is a known calcium binding protein and an efficient calcium buffer. Therefore, in the present study we hypothesized that downregulation of Dd-Grp94 protein would affect Dictyostelium cell structure, growth, and development. We found that Dd-grp94 RNAi recombinants exhibited reduced growth rate, cell size, and a subtle change in cell motility compared to the parental cells. The recombinants also exhibited a delay in development and small fruiting bodies. These results establish that Dd-grp94 plays a crucial role in determining normal cell structure, growth and differentiation.

Caffeine sensitive repair and mutation induction in UV- or γ-ray-irradiated Dictyostelium discoideum

International Nuclear Information System (INIS)

Kanishi, Nobuji; Kinjo, Yasuhito; Watanabe, Makoto.

1990-01-01

It seems that certain kinds of chemical substances increase the distortion in molecules, change the high order microstructures of nuclei and chromosomes, and exert large variation to the function of repairing the damage of genes due to radiation and others, by coupling with DNA, protein or enzyme system. It has been well known that caffeine is one of such compounds, and by coupling with DNA, it increases the damage due to ultraviolet ray and gives the action of obstructing repair in addition to the action of inducing the abnormality of chromosomes and mutation. Dictyostelium discoideum has the simplest nuclear structure, and shows extremely high resistance to radiation by its high restoration ability. The authors have advanced the research by paying attention to its characteristics, and comparing the Dictyostelium discoideum as one model system with the lymphocyte system of higher animals. This time, the authors analyzed the characteristics of two kinds of sensitivity repair process of caffeine, and investigated into their relation with the occurrence of mutation. The experimental method and the results are reported. (K.I.)

Caffeine sensitive repair and mutation induction in UV- or. gamma. -ray-irradiated Dictyostelium discoideum

Energy Technology Data Exchange (ETDEWEB)

Kanishi, Nobuji (Tokyo Metropolitan Research Lab. of Public Health (Japan)); Kinjo, Yasuhito; Watanabe, Makoto

1990-01-01

It seems that certain kinds of chemical substances increase the distortion in molecules, change the high order microstructures of nuclei and chromosomes, and exert large variation to the function of repairing the damage of genes due to radiation and others, by coupling with DNA, protein or enzyme system. It has been well known that caffeine is one of such compounds, and by coupling with DNA, it increases the damage due to ultraviolet ray and gives the action of obstructing repair in addition to the action of inducing the abnormality of chromosomes and mutation. Dictyostelium discoideum has the simplest nuclear structure, and shows extremely high resistance to radiation by its high restoration ability. The authors have advanced the research by paying attention to its characteristics, and comparing the Dictyostelium discoideum as one model system with the lymphocyte system of higher animals. This time, the authors analyzed the characteristics of two kinds of sensitivity repair process of caffeine, and investigated into their relation with the occurrence of mutation. The experimental method and the results are reported. (K.I.).

A secreted protein is an endogenous chemorepellant in Dictyostelium discoideum.

Science.gov (United States)

Phillips, Jonathan E; Gomer, Richard H

2012-07-03

Chemorepellants may play multiple roles in physiological and pathological processes. However, few endogenous chemorepellants have been identified, and how they function is unclear. We found that the autocrine signal AprA, which is produced by growing Dictyostelium discoideum cells and inhibits their proliferation, also functions as a chemorepellant. Wild-type cells at the edge of a colony show directed movement outward from the colony, whereas cells lacking AprA do not. Cells show directed movement away from a source of recombinant AprA and dialyzed conditioned media from wild-type cells, but not dialyzed conditioned media from aprA(-) cells. The secreted protein CfaD, the G protein Gα8, and the kinase QkgA are necessary for the chemorepellant activity of AprA as well as its proliferation-inhibiting activity, whereas the putative transcription factor BzpN is dispensable for the chemorepellant activity of AprA but necessary for inhibition of proliferation. Phospholipase C and PI3 kinases 1 and 2, which are necessary for the activity of at least one other chemorepellant in Dictyostelium, are not necessary for recombinant AprA chemorepellant activity. Starved cells are not repelled by recombinant AprA, suggesting that aggregation-phase cells are not sensitive to the chemorepellant effect. Cell tracking indicates that AprA affects the directional bias of cell movement, but not cell velocity or the persistence of cell movement. Together, our data indicate that the endogenous signal AprA acts as an autocrine chemorepellant for Dictyostelium cells.

Immobilization of Deoxyadenosine Kinase from Dictyostelium discoideum (DddAK) and Its Application in the 5’-Phosphorylation of Arabinosyladenine and Arabinosyl-2-fluoroadenine

DEFF Research Database (Denmark)

Serra, Immacolata; Ubiali, Daniela; Piskur, Jure

2017-01-01

Deoxyadenosine kinase from Dictyostelium discoideum (DddAK) phosphorylates its natural substrate (2’‐deoxyadenosine, dAdo) as well as the arabinosyladenine analogues vidarabine (araA) and fludarabine (F‐araA) to their corresponding 5’‐monophosphates. DddAK has been here immobilized by ionic...... interaction on an aminated epoxy‐functionalized support (SepabeadsTM EC‐EP), and cross‐linked with oxidized dextran. The final activity recovery was 33–42 %, depending on the protein loading. Immobilization enhanced the stability of DddAK at pH 10 and, to a lesser extent, at 45 °C. Phosphorylation of d...

Cytoplasmic pH and the regulation of the dictyostelium cell cycle

NARCIS (Netherlands)

Aerts, R.J.; Durston, A.J.; Moolenaar, W.H.

1985-01-01

Cytoplasmic pH (pHl) was monitored during the cell cycle of synchronous populations of Dictyostelium discoideum by means of a pH “null point” method. There is a cycle of pHl that closely corresponds to the DNA replication cycle, with a minimum of pH 7.20 in interphase and a peak of pH 7.45 during S

The Dictyostelium discoideum cellulose synthase: Structure/function analysis and identification of interacting proteins

Energy Technology Data Exchange (ETDEWEB)

Richard L. Blanton

2004-02-19

OAK-B135 The major accomplishments of this project were: (1) the initial characterization of dcsA, the gene for the putative catalytic subunit of cellulose synthase in the cellular slime mold Dictyostelium discoideum; (2) the detection of a developmentally regulated event (unidentified, but perhaps a protein modification or association with a protein partner) that is required for cellulose synthase activity (i.e., the dcsA product is necessary, but not sufficient for cellulose synthesis); (3) the continued exploration of the developmental context of cellulose synthesis and DcsA; (4) the isolation of a GFP-DcsA-expressing strain (work in progress); and (5) the identification of Dictyostelium homologues for plant genes whose products play roles in cellulose biosynthesis. Although our progress was slow and many of our results negative, we did develop a number of promising avenues of investigation that can serve as the foundation for future projects.

Xpf and not the Fanconi anaemia proteins or Rev3 accounts for the extreme resistance to cisplatin in Dictyostelium discoideum.

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Xiao-Yin Zhang

2009-09-01

Full Text Available Organisms like Dictyostelium discoideum, often referred to as DNA damage "extremophiles", can survive exposure to extremely high doses of radiation and DNA crosslinking agents. These agents form highly toxic DNA crosslinks that cause extensive DNA damage. However, little is known about how Dictyostelium and the other "extremophiles" can tolerate and repair such large numbers of DNA crosslinks. Here we describe a comprehensive genetic analysis of crosslink repair in Dictyostelium discoideum. We analyse three gene groups that are crucial for a replication-coupled repair process that removes DNA crosslinks in higher eukarya: The Fanconi anaemia pathway (FA, translesion synthesis (TLS, and nucleotide excision repair. Gene disruption studies unexpectedly reveal that the FA genes and the TLS enzyme Rev3 play minor roles in tolerance to crosslinks in Dictyostelium. However, disruption of the Xpf nuclease subcomponent results in striking hypersensitivity to crosslinks. Genetic interaction studies reveal that although Xpf functions with FA and TLS gene products, most Xpf mediated repair is independent of these two gene groups. These results suggest that Dictyostelium utilises a distinct Xpf nuclease-mediated repair process to remove crosslinked DNA. Other DNA damage-resistant organisms and chemoresistant cancer cells might adopt a similar strategy to develop resistance to DNA crosslinking agents.

An Autocrine Proliferation Repressor Regulates Dictyostelium discoideum Proliferation and Chemorepulsion Using the G Protein-Coupled Receptor GrlH

OpenAIRE

Yu Tang; Yuantai Wu; Sarah E. Herlihy; Francisco J. Brito-Aleman; Jose H. Ting; Chris Janetopoulos; Richard H. Gomer; Scott D. Emr

2018-01-01

In eukaryotic microbes, little is known about signals that inhibit the proliferation of the cells that secrete the signal, and little is known about signals (chemorepellents) that cause cells to move away from the source of the signal. Autocrine proliferation repressor protein A (AprA) is a protein secreted by the eukaryotic microbe Dictyostelium discoideum. AprA is a chemorepellent for and inhibits the proliferation of D. discoideum. We previously found that cells sense AprA using G proteins...

Binding and assembly of actin filaments by plasma membranes from dictyostelium discoideum

International Nuclear Information System (INIS)

Schwartz, M.A.; Luna, E.J.

1986-01-01

The binding of native, 125 I-Bolton-Hunter-labeled actin to purified Dictyostelium discoideum plasma membranes was measured using a sedimentation assay. Binding was saturable only in the presence of the actin capping protein, gelsolin. The binding curves were sigmoidal, indicating positive cooperativity at low actin concentrations. This cooperativity appeared to be due to actin-actin associations during polymerization, since phalloidin converted the curve to a hyperbolic shape. This membrane-bound actin stained with rhodamine-phalloidin and was cross-linked by m-maleimidobenzoyl succinimide ester, a bifunctional cross-linker, into multimers with the same pattern observed for cross-linked F-actin. The authors conclude that D. discoideum plasma membranes bind actin specifically and saturably and that these membranes organize actin into filaments below the normal critical concentration for polymerization. This interaction probably occurs between multiple binding sites on the membrane and the side of the actin filament, and may be related to the clustering of membrane proteins

Influence of fast advective flows on pattern formation of Dictyostelium discoideum

Science.gov (United States)

Bae, Albert; Zykov, Vladimir; Bodenschatz, Eberhard

2018-01-01

We report experimental and numerical results on pattern formation of self-organizing Dictyostelium discoideum cells in a microfluidic setup under a constant buffer flow. The external flow advects the signaling molecule cyclic adenosine monophosphate (cAMP) downstream, while the chemotactic cells attached to the solid substrate are not transported with the flow. At high flow velocities, elongated cAMP waves are formed that cover the whole length of the channel and propagate both parallel and perpendicular to the flow direction. While the wave period and transverse propagation velocity are constant, parallel wave velocity and the wave width increase linearly with the imposed flow. We also observe that the acquired wave shape is highly dependent on the wave generation site and the strength of the imposed flow. We compared the wave shape and velocity with numerical simulations performed using a reaction-diffusion model and found excellent agreement. These results are expected to play an important role in understanding the process of pattern formation and aggregation of D. discoideum that may experience fluid flows in its natural habitat. PMID:29590179

Identification of Pentatricopeptide Repeat Proteins in the Model Organism Dictyostelium discoideum

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Sam Manna

2013-01-01

Full Text Available Pentatricopeptide repeat (PPR proteins are RNA binding proteins with functions in organelle RNA metabolism. They are found in all eukaryotes but have been most extensively studied in plants. We report on the identification of 12 PPR-encoding genes in the genome of the protist Dictyostelium discoideum, with potential homologs in other members of the same lineage and some predicted novel functions for the encoded gene products in protists. For one of the gene products, we show that it localizes to the mitochondria, and we also demonstrate that antisense inhibition of its expression leads to slower growth, a phenotype associated with mitochondrial dysfunction.

Regulation of TORC2 complex in Dictyostelium discoideum

NARCIS (Netherlands)

Khanna, Ankita

2016-01-01

Dictyostelium is an amoeba that lives in the soil where it feeds on bacteria. During scarcity of food, Dictyostelium cells undergo a highly regulated developmental process in which the cells aggregate by chemotaxing towards pulsatile emission of extracellular cAMP from a signaling center; the cells

A new social gene in Dictyostelium discoideum, chtB

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Santorelli Lorenzo A

2013-01-01

Full Text Available Abstract Background Competitive social interactions are ubiquitous in nature, but their genetic basis is difficult to determine. Much can be learned from single gene knockouts in a eukaryote microbe. The mutants can be competed with the parent to discern the social impact of that specific gene. Dictyostelium discoideum is a social amoeba that exhibits cooperative behavior in the construction of a multicellular fruiting body. It is a good model organism to study the genetic basis of cooperation since it has a sequenced genome and it is amenable to genetic manipulation. When two strains of D. discoideum are mixed, a cheater strain can exploit its social partner by differentiating more spore than its fair share relative to stalk cells. Cheater strains can be generated in the lab or found in the wild and genetic analyses have shown that cheating behavior can be achieved through many pathways. Results We have characterized the knockout mutant chtB, which was isolated from a screen for cheater mutants that were also able to form normal fruiting bodies on their own. When mixed in equal proportions with parental strain cells, chtB mutants contributed almost 60% of the total number of spores. To do so, chtB cells inhibit wild type cells from becoming spores, as indicated by counts and by the wild type cells’ reduced expression of the prespore gene, cotB. We found no obvious fitness costs (morphology, doubling time in liquid medium, spore production, and germination efficiency associated with the cheating ability of the chtB knockout. Conclusions In this study we describe a new gene in D. discoideum, chtB, which when knocked out inhibits the parental strain from producing spores. Moreover, under lab conditions, we did not detect any fitness costs associated with this behavior.

Yersinia outer protein YopE affects the actin cytoskeleton in Dictyostelium discoideum through targeting of multiple Rho family GTPases

LENUS (Irish Health Repository)

Vlahou, Georgia

2009-07-14

Abstract Background All human pathogenic Yersinia species share a virulence-associated type III secretion system that translocates Yersinia effector proteins into host cells to counteract infection-induced signaling responses and prevent phagocytosis. Dictyostelium discoideum has been recently used to study the effects of bacterial virulence factors produced by internalized pathogens. In this study we explored the potential of Dictyostelium as model organism for analyzing the effects of ectopically expressed Yersinia outer proteins (Yops). Results The Yersinia pseudotuberculosis virulence factors YopE, YopH, YopM and YopJ were expressed de novo within Dictyostelium and their effects on growth in axenic medium and on bacterial lawns were analyzed. No severe effect was observed for YopH, YopJ and YopM, but expression of YopE, which is a GTPase activating protein for Rho GTPases, was found to be highly detrimental. GFP-tagged YopE expressing cells had less conspicuous cortical actin accumulation and decreased amounts of F-actin. The actin polymerization response upon cAMP stimulation was impaired, although chemotaxis was unaffected. YopE also caused reduced uptake of yeast particles. These alterations are probably due to impaired Rac1 activation. We also found that YopE predominantly associates with intracellular membranes including the Golgi apparatus and inhibits the function of moderately overexpressed RacH. Conclusion The phenotype elicited by YopE in Dictyostelium can be explained, at least in part, by inactivation of one or more Rho family GTPases. It further demonstrates that the social amoeba Dictyostelium discoideum can be used as an efficient and easy-to-handle model organism in order to analyze the function of a translocated GAP protein of a human pathogen.

Subcellular localization of ammonium transporters in Dictyostelium discoideum

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Davis Carter T

2008-12-01

Full Text Available Abstract Background With the exception of vertebrates, most organisms have plasma membrane associated ammonium transporters which primarily serve to import a source of nitrogen for nutritional purposes. Dictyostelium discoideum has three ammonium transporters, Amts A, B and C. Our present work used fluorescent fusion proteins to determine the cellular localization of the Amts and tested the hypothesis that the transporters mediate removal of ammonia generated endogenously from the elevated protein catabolism common to many protists. Results Using RFP and YFP fusion constructs driven by the actin 15 promoter, we found that the three ammonium transporters were localized on the plasma membrane and on the membranes of subcellular organelles. AmtA and AmtB were localized on the membranes of endolysosomes and phagosomes, with AmtB further localized on the membranes of contractile vacuoles. AmtC also was localized on subcellular organelles when it was stabilized by coexpression with either the AmtA or AmtB fusion transporter. The three ammonium transporters exported ammonia linearly with regard to time during the first 18 hours of the developmental program as revealed by reduced export in the null strains. The fluorescently tagged transporters rescued export when expressed in the null strains, and thus they were functional transporters. Conclusion Unlike ammonium transporters in most organisms, which import NH3/NH4+ as a nitrogen source, those of Dictyostelium export ammonia/ammonium as a waste product from extensive catabolism of exogenously derived and endogenous proteins. Localization on proteolytic organelles and on the neutral contractile vacuole suggests that Dictyostelium ammonium transporters may have unique subcellular functions and play a role in the maintenance of intracellular ammonium distribution. A lack of correlation between the null strain phenotypes and ammonia excretion properties of the ammonium transporters suggests that it is not

Specificity of the Cyclic GMP-Binding Activity and of a Cyclic GMP-Dependent Cyclic GMP Phosphodiesterase in Dictyostelium discoideum

NARCIS (Netherlands)

Haastert, Peter J.M. van; Walsum, Hans van; Meer, Rob C. van der; Bulgakov, Roman; Konijn, Theo M.

1982-01-01

The nucleotide specificity of the cyclic GMP-binding activity in a homogenate of Dictyostelium discoideum was determined by competition of cyclic GMP derivatives with [8-3H] cyclic GMP for the binding sites. The results indicate that cyclic GMP is bound to the binding proteins by hydrogen bonds at

Structural and functional studies of a family of Dictyostelium discoideum developmentally regulated, prestalk genes coding for small proteins

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Escalante Ricardo

2008-01-01

Full Text Available Abstract Background The social amoeba Dictyostelium discoideum executes a multicellular development program upon starvation. This morphogenetic process requires the differential regulation of a large number of genes and is coordinated by extracellular signals. The MADS-box transcription factor SrfA is required for several stages of development, including slug migration and spore terminal differentiation. Results Subtractive hybridization allowed the isolation of a gene, sigN (SrfA-induced gene N, that was dependent on the transcription factor SrfA for expression at the slug stage of development. Homology searches detected the existence of a large family of sigN-related genes in the Dictyostelium discoideum genome. The 13 most similar genes are grouped in two regions of chromosome 2 and have been named Group1 and Group2 sigN genes. The putative encoded proteins are 87–89 amino acids long. All these genes have a similar structure, composed of a first exon containing a 13 nucleotides long open reading frame and a second exon comprising the remaining of the putative coding region. The expression of these genes is induced at10 hours of development. Analyses of their promoter regions indicate that these genes are expressed in the prestalk region of developing structures. The addition of antibodies raised against SigN Group 2 proteins induced disintegration of multi-cellular structures at the mound stage of development. Conclusion A large family of genes coding for small proteins has been identified in D. discoideum. Two groups of very similar genes from this family have been shown to be specifically expressed in prestalk cells during development. Functional studies using antibodies raised against Group 2 SigN proteins indicate that these genes could play a role during multicellular development.

Cheating by exploitation of developmental prestalk patterning in Dictyostelium discoideum.

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Anupama Khare

2010-02-01

Full Text Available The cooperative developmental system of the social amoeba Dictyostelium discoideum is susceptible to exploitation by cheaters-strains that make more than their fair share of spores in chimerae. Laboratory screens in Dictyostelium have shown that the genetic potential for facultative cheating is high, and field surveys have shown that cheaters are abundant in nature, but the cheating mechanisms are largely unknown. Here we describe cheater C (chtC, a strong facultative cheater mutant that cheats by affecting prestalk differentiation. The chtC gene is developmentally regulated and its mRNA becomes stalk-enriched at the end of development. chtC mutants are defective in maintaining the prestalk cell fate as some of their prestalk cells transdifferentiate into prespore cells, but that defect does not affect gross developmental morphology or sporulation efficiency. In chimerae between wild-type and chtC mutant cells, the wild-type cells preferentially give rise to prestalk cells, and the chtC mutants increase their representation in the spore mass. Mixing chtC mutants with other cell-type proportioning mutants revealed that the cheating is directly related to the prestalk-differentiation propensity of the victim. These findings illustrate that a cheater can victimize cooperative strains by exploiting an established developmental pathway.

Cheating by Exploitation of Developmental Prestalk Patterning in Dictyostelium discoideum

Science.gov (United States)

Khare, Anupama; Shaulsky, Gad

2010-01-01

The cooperative developmental system of the social amoeba Dictyostelium discoideum is susceptible to exploitation by cheaters—strains that make more than their fair share of spores in chimerae. Laboratory screens in Dictyostelium have shown that the genetic potential for facultative cheating is high, and field surveys have shown that cheaters are abundant in nature, but the cheating mechanisms are largely unknown. Here we describe cheater C (chtC), a strong facultative cheater mutant that cheats by affecting prestalk differentiation. The chtC gene is developmentally regulated and its mRNA becomes stalk-enriched at the end of development. chtC mutants are defective in maintaining the prestalk cell fate as some of their prestalk cells transdifferentiate into prespore cells, but that defect does not affect gross developmental morphology or sporulation efficiency. In chimerae between wild-type and chtC mutant cells, the wild-type cells preferentially give rise to prestalk cells, and the chtC mutants increase their representation in the spore mass. Mixing chtC mutants with other cell-type proportioning mutants revealed that the cheating is directly related to the prestalk-differentiation propensity of the victim. These findings illustrate that a cheater can victimize cooperative strains by exploiting an established developmental pathway. PMID:20195510

Characterization of a 1,4-. beta. -D-glucan synthase from Dictyostelium discoideum

Energy Technology Data Exchange (ETDEWEB)

Blanton, R.L.

1992-01-15

Various aspects of research concerning Dictyostelium discoideum are presented. The initial focus of this project was upon: the characterization of potential probes for the cellulose synthase (antibody and nucleic acid), the determination of the cultural induction conditions of cellulose synthesis, the solubilization of the enzyme activity, the development of a non-inhibitory disruption buffer, the generation and isolation of mutant strains deficient in cellulose synthesis, and the development of the capability to determine the degree of polymerization of the in vitro product. I have briefly summarized our most significant findings with only selected data sets being shown in this report in the interest of brevity.

Variation, sex, and social cooperation: molecular population genetics of the social amoeba Dictyostelium discoideum.

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Jonathan M Flowers

2010-07-01

Full Text Available Dictyostelium discoideum is a eukaryotic microbial model system for multicellular development, cell-cell signaling, and social behavior. Key models of social evolution require an understanding of genetic relationships between individuals across the genome or possibly at specific genes, but the nature of variation within D. discoideum is largely unknown. We re-sequenced 137 gene fragments in wild North American strains of D. discoideum and examined the levels and patterns of nucleotide variation in this social microbial species. We observe surprisingly low levels of nucleotide variation in D. discoideum across these strains, with a mean nucleotide diversity (pi of 0.08%, and no strong population stratification among North American strains. We also do not find any clear relationship between nucleotide divergence between strains and levels of social dominance and kin discrimination. Kin discrimination experiments, however, show that strains collected from the same location show greater ability to distinguish self from non-self than do strains from different geographic areas. This suggests that a greater ability to recognize self versus non-self may arise among strains that are more likely to encounter each other in nature, which would lead to preferential formation of fruiting bodies with clonemates and may prevent the evolution of cheating behaviors within D. discoideum populations. Finally, despite the fact that sex has rarely been observed in this species, we document a rapid decay of linkage disequilibrium between SNPs, the presence of recombinant genotypes among natural strains, and high estimates of the population recombination parameter rho. The SNP data indicate that recombination is widespread within D. discoideum and that sex as a form of social interaction is likely to be an important aspect of the life cycle.

Mitochondrial Stress Tests Using Seahorse Respirometry on Intact Dictyostelium discoideum Cells.

Science.gov (United States)

Lay, Sui; Sanislav, Oana; Annesley, Sarah J; Fisher, Paul R

2016-01-01

Mitochondria not only play a critical and central role in providing metabolic energy to the cell but are also integral to the other cellular processes such as modulation of various signaling pathways. These pathways affect many aspects of cell physiology, including cell movement, growth, division, differentiation, and death. Mitochondrial dysfunction which affects mitochondrial bioenergetics and causes oxidative phosphorylation defects can thus lead to altered cellular physiology and manifest in disease. The assessment of the mitochondrial bioenergetics can thus provide valuable insights into the physiological state, and the alterations to the state of the cells. Here, we describe a method to successfully use the Seahorse XF(e)24 Extracellular Flux Analyzer to assess the mitochondrial respirometry of the cellular slime mold Dictyostelium discoideum.

Identification and characterization of peptide: N- glycanase from Dictyostelium discoideum

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Gosain Anuradha

2012-06-01

Full Text Available Abstract Background Peptide: N- glycanase (PNGase enzyme cleaves oligosaccharides from the misfolded glycoproteins and prepares them for degradation. This enzyme plays a role in the endoplasmic reticulum associated degradation (ERAD pathway in yeast and mice but its biological importance and role in multicellular development remain largely unknown. Results In this study, the PNGase from the cellular slime mold, Dictyostelium discoideum (DdPNGase was identified based on the presence of a common TG (transglutaminase core domain and its sequence homology with the known PNGases. The domain architecture and the sequence comparison validated the presence of probable functional domains in DdPNGase. The tertiary structure matched with the mouse PNGase. Here we show that DdPNGase is an essential protein, required for aggregation during multicellular development and a knockout strain of it results in small sized aggregates, all of which did not form fruiting bodies. The in situ hybridization and RT-PCR results show higher level of expression during the aggregate stage. The expression gets restricted to the prestalk region during later developmental stages. DdPNGase is a functional peptide:N-glycanase enzyme possessing deglycosylation activity, but does not possess any significant transamidation activity. Conclusions We have identified and characterized a novel PNGase from D. discoideum and confirmed its deglycosylation activity. The results emphasize the importance of PNGase in aggregation during multicellular development of this organism.

Sociogenomics of self vs. non-self cooperation during development of Dictyostelium discoideum.

Science.gov (United States)

Li, Si I; Buttery, Neil J; Thompson, Christopher R L; Purugganan, Michael D

2014-07-21

Dictyostelium discoideum, a microbial model for social evolution, is known to distinguish self from non-self and show genotype-dependent behavior during chimeric development. Aside from a small number of cell-cell recognition genes, however, little is known about the genetic basis of self/non-self recognition in this species. Based on the key hypothesis that there should be differential expression of genes if D. discoideum cells were interacting with non-clone mates, we performed transcriptomic profiling study in this species during clonal vs. chimeric development. The transcriptomic profiles of D. discoideum cells in clones vs. different chimeras were compared at five different developmental stages using a customized microarray. Effects of chimerism on global transcriptional patterns associated with social interactions were observed. We find 1,759 genes significantly different between chimera and clone, 1,144 genes associated significant strain differences, and 6,586 genes developmentally regulated over time. Principal component analysis showed a small amount of the transcriptional variance to chimerism-related factors (Chimerism: 0.18%, Chimerism × Timepoint: 0.03%). There are 162 genes specifically regulated under chimeric development, with continuous small differences between chimera vs. clone over development. Almost 60% of chimera-associated differential genes were differentially expressed at the 4 h aggregate stage, which corresponds to the initial transition of D. discoideum from solitary life to a multicellular phase. A relatively small proportion of over-all variation in gene expression is explained by differences between chimeric and clonal development. The relatively small modifications in gene expression associated with chimerism is compatible with the high level of cooperation observed among different strains of D. discoideum; cells of distinct genetic backgrounds will co-aggregate indiscriminately and co-develop into fruiting bodies. Chimeric

Functional characterisation of parvulin-type peptidyl prolyl cis-trans isomerase, PinA in Dictyostelium discoideum

International Nuclear Information System (INIS)

Haokip, Nemneineng; Naorem, Aruna

2017-01-01

Pin1-type parvulins are unique among PPIases that can catalyse an otherwise slow cis-trans isomerisation of phosphorylated peptide bond preceding proline in target proteins. This prolyl isomerisation process can regulate activity, stability and localisation of target proteins and thus control cellular processes like eukaryotic cell proliferation, cell cycle progression and gene regulation. Towards understanding the function of Pin1-type prolyl isomerisation in Dictyostelium discoideum, a slime mould with distinct growth and developmental phases, we identified PinA as a novel Pin1-type parvulin by its ability to complement the temperature sensitivity phenotype associated with a mutation in ESS1 in S. cerevisiae. In D. discoideum, pinA is temporally and spatially regulated during growth and development. PinA is both nuclear as well as cytoplasmic in the growing cells. We further show that loss of pinA (pinA − ) leads to decreased growth rate, reduced spore formation and abnormal prespore-prestalk patterning. We conclude that PinA is required for normal growth as well as development in D. discoideum. - Highlights: • PinA is a bona fide homologue of S. cerevisiae Ess1. • PinA is required for normal cell proliferation of D. discoideum. • PinA is spatially localised in developmental structures. • PinA is important for cell differentiation and patterning.

Nucleocytoplasmic protein translocation during mitosis in the social amoebozoan Dictyostelium discoideum.

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O'Day, Danton H; Budniak, Aldona

2015-02-01

Mitosis is a fundamental and essential life process. It underlies the duplication and survival of all cells and, as a result, all eukaryotic organisms. Since uncontrolled mitosis is a dreaded component of many cancers, a full understanding of the process is critical. Evolution has led to the existence of three types of mitosis: closed, open, and semi-open. The significance of these different mitotic species, how they can lead to a full understanding of the critical events that underlie the asexual duplication of all cells, and how they may generate new insights into controlling unregulated cell division remains to be determined. The eukaryotic microbe Dictyostelium discoideum has proved to be a valuable biomedical model organism. While it appears to utilize closed mitosis, a review of the literature suggests that it possesses a form of mitosis that lies in the middle between truly open and fully closed mitosis-it utilizes a form of semi-open mitosis. Here, the nucleocytoplasmic translocation patterns of the proteins that have been studied during mitosis in the social amoebozoan D. discoideum are detailed followed by a discussion of how some of them provide support for the hypothesis of semi-open mitosis. © 2014 The Authors. Biological Reviews published by John Wiley & Sons Ltd on behalf of Cambridge Philosophical Society.

Effects of Nickel, Chlorpyrifos and Their Mixture on the Dictyostelium discoideum Proteome

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Boatti, Lara; Robotti, Elisa; Marengo, Emilio; Viarengo, Aldo; Marsano, Francesco

2012-01-01

Mixtures of chemicals can have additive, synergistic or antagonistic interactions. We investigated the effects of the exposure to nickel, the organophosphate insecticide chlorpyrifos at effect concentrations (EC) of 25% and 50% and their binary mixture (Ec25 + EC25) on Dictyostelium discoideum amoebae based on lysosomal membrane stability (LMS). We treated D. discoideum with these compounds under controlled laboratory conditions and evaluated the changes in protein levels using a two-dimensional gel electrophoresis (2DE) proteomic approach. Nickel treatment at EC25 induced changes in 14 protein spots, 12 of which were down-regulated. Treatment with nickel at EC50 resulted in changes in 15 spots, 10 of which were down-regulated. Treatment with chlorpyrifos at EC25 induced changes in six spots, all of which were down-regulated; treatment with chlorpyrifos at EC50 induced changes in 13 spots, five of which were down-regulated. The mixture corresponding to EC25 of each compound induced changes in 19 spots, 13 of which were down-regulated. The data together reveal that a different protein expression signature exists for each treatment, and that only a few proteins are modulated in multiple different treatments. For a simple binary mixture, the proteomic response does not allow for the identification of each toxicant. The protein spots that showed significant differences were identified by mass spectrometry, which revealed modulations of proteins involved in metal detoxification, stress adaptation, the oxidative stress response and other cellular processes. PMID:23443088

Transcriptional down-regulation and rRNA cleavage in Dictyostelium discoideum mitochondria during Legionella pneumophila infection.

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Chenyu Zhang

2009-05-01

Full Text Available Bacterial pathogens employ a variety of survival strategies when they invade eukaryotic cells. The amoeba Dictyostelium discoideum is used as a model host to study the pathogenic mechanisms that Legionella pneumophila, the causative agent of Legionnaire's disease, uses to kill eukaryotic cells. Here we show that the infection of D. discoideum by L. pneumophila results in a decrease in mitochondrial messenger RNAs, beginning more than 8 hours prior to detectable host cell death. These changes can be mimicked by hydrogen peroxide treatment, but not by other cytotoxic agents. The mitochondrial large subunit ribosomal RNA (LSU rRNA is also cleaved at three specific sites during the course of infection. Two LSU rRNA fragments appear first, followed by smaller fragments produced by additional cleavage events. The initial LSU rRNA cleavage site is predicted to be on the surface of the large subunit of the mitochondrial ribosome, while two secondary sites map to the predicted interface with the small subunit. No LSU rRNA cleavage was observed after exposure of D. discoideum to hydrogen peroxide, or other cytotoxic chemicals that kill cells in a variety of ways. Functional L. pneumophila type II and type IV secretion systems are required for the cleavage, establishing a correlation between the pathogenesis of L. pneumophila and D. discoideum LSU rRNA destruction. LSU rRNA cleavage was not observed in L. pneumophila infections of Acanthamoeba castellanii or human U937 cells, suggesting that L. pneumophila uses distinct mechanisms to interrupt metabolism in different hosts. Thus, L. pneumophila infection of D. discoideum results in dramatic decrease of mitochondrial RNAs, and in the specific cleavage of mitochondrial rRNA. The predicted location of the cleavage sites on the mitochondrial ribosome suggests that rRNA destruction is initiated by a specific sequence of events. These findings suggest that L. pneumophila specifically disrupts mitochondrial

Effect of sodium fluoride on the amount of polyribosomes, single ribosomes and ribosomal subunits in a cellular slime mold, Dictyostelium discoideum

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Sameshima, M; Ito, K; Iwabuchi, M

1972-01-01

In the slime mold, Dictyostelium discoideum, when the rate of protein synthesis was decreased by NaF, free 80-S ribosomes accumulated at the expense of polyribosomes, while 60-S and 40-S ribosomal subunits remained almost constant. The same level of ribosomal subunits was also maintained in cells after incubation with cycloheximide or at the stationary phase of growth.

Whole genome sequencing of mutation accumulation lines reveals a low mutation rate in the social amoeba Dictyostelium discoideum.

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Gerda Saxer

Full Text Available Spontaneous mutations play a central role in evolution. Despite their importance, mutation rates are some of the most elusive parameters to measure in evolutionary biology. The combination of mutation accumulation (MA experiments and whole-genome sequencing now makes it possible to estimate mutation rates by directly observing new mutations at the molecular level across the whole genome. We performed an MA experiment with the social amoeba Dictyostelium discoideum and sequenced the genomes of three randomly chosen lines using high-throughput sequencing to estimate the spontaneous mutation rate in this model organism. The mitochondrial mutation rate of 6.76×10(-9, with a Poisson confidence interval of 4.1×10(-9 - 9.5×10(-9, per nucleotide per generation is slightly lower than estimates for other taxa. The mutation rate estimate for the nuclear DNA of 2.9×10(-11, with a Poisson confidence interval ranging from 7.4×10(-13 to 1.6×10(-10, is the lowest reported for any eukaryote. These results are consistent with low microsatellite mutation rates previously observed in D. discoideum and low levels of genetic variation observed in wild D. discoideum populations. In addition, D. discoideum has been shown to be quite resistant to DNA damage, which suggests an efficient DNA-repair mechanism that could be an adaptation to life in soil and frequent exposure to intracellular and extracellular mutagenic compounds. The social aspect of the life cycle of D. discoideum and a large portion of the genome under relaxed selection during vegetative growth could also select for a low mutation rate. This hypothesis is supported by a significantly lower mutation rate per cell division in multicellular eukaryotes compared with unicellular eukaryotes.

An Autocrine Proliferation Repressor Regulates Dictyostelium discoideum Proliferation and Chemorepulsion Using the G Protein-Coupled Receptor GrlH

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Yu Tang

2018-02-01

Full Text Available In eukaryotic microbes, little is known about signals that inhibit the proliferation of the cells that secrete the signal, and little is known about signals (chemorepellents that cause cells to move away from the source of the signal. Autocrine proliferation repressor protein A (AprA is a protein secreted by the eukaryotic microbe Dictyostelium discoideum. AprA is a chemorepellent for and inhibits the proliferation of D. discoideum. We previously found that cells sense AprA using G proteins, suggesting the existence of a G protein-coupled AprA receptor. To identify the AprA receptor, we screened mutants lacking putative G protein-coupled receptors. We found that, compared to the wild-type strain, cells lacking putative receptor GrlH (grlH{macron} cells show rapid proliferation, do not have large numbers of cells moving away from the edges of colonies, are insensitive to AprA-induced proliferation inhibition and chemorepulsion, and have decreased AprA binding. Expression of GrlH in grlH{macron} cells (grlH{macron}/grlHOE rescues the phenotypes described above. These data indicate that AprA signaling may be mediated by GrlH in D. discoideum.

Down-regulation of Cell Surface Cyclic AMP Receptors and Desensitization of Cyclic AMP-stimulated Adenylate Cyclase by Cyclic AMP in Dictyostelium discoideum. Kinetics and Concentration Dependence

NARCIS (Netherlands)

Haastert, Peter J.M. van

1987-01-01

cAMP binds to Dictyostelium discoideum surface receptors and induces a transient activation of adenylate cyclase, which is followed by desensitization. cAMP also induces a loss of detectable surface receptors (down-regulation). Cells were incubated with constant cAMP concentrations, washed free of

A retinoblastoma orthologue is required for the sensing of a chalone in Dictyostelium discoideum.

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Bakthavatsalam, Deenadayalan; White, Michael J V; Herlihy, Sarah E; Phillips, Jonathan E; Gomer, Richard H

2014-03-01

Retinoblastoma-like proteins regulate cell differentiation and inhibit cell proliferation. The Dictyostelium discoideum retinoblastoma orthologue RblA affects the differentiation of cells during multicellular development, but it is unclear whether RblA has a significant effect on Dictyostelium cell proliferation, which is inhibited by the secreted proteins AprA and CfaD. We found that rblA⁻ cells in shaking culture proliferate to a higher density, die faster after reaching stationary density, and, after starvation, have a lower spore viability than wild-type cells, possibly because in shaking culture, rblA⁻ cells have both increased cytokinesis and lower extracellular accumulation of CfaD. However, rblA⁻ cells have abnormally slow proliferation on bacterial lawns. Recombinant AprA inhibits the proliferation of wild-type cells but not that of rblA⁻ cells, whereas CfaD inhibits the proliferation of both wild-type cells and rblA⁻ cells. Similar to aprA⁻ cells, rblA⁻ cells have a normal mass and protein accumulation rate on a per-nucleus basis, indicating that RblA affects cell proliferation but not cell growth. AprA also functions as a chemorepellent, and RblA is required for proper AprA chemorepellent activity despite the fact that RblA does not affect cell speed. Together, our data indicate that an autocrine proliferation-inhibiting factor acts through RblA to regulate cell density in Dictyostelium, suggesting that such factors may signal through retinoblastoma-like proteins to control the sizes of structures such as developing organs or tumors.

Curcumin inhibits development and cell adhesion in Dictyostelium discoideum: Implications for YakA signaling and GST enzyme function

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Garige, Mamatha; Walters, Eric, E-mail: ewalters@howard.edu

2015-11-13

The molecular basis for nutraceutical properties of the polyphenol curcumin (Curcuma longa, Turmeric) is complex, affecting multiple factors that regulate cell signaling and homeostasis. Here, we report the effect of curcumin on cellular and developmental mechanisms in the eukaryotic model, Dictyostelium discoideum. Dictyostelium proliferation was inhibited in the presence of curcumin, which also suppressed the prestarvation marker, discoidin I, members of the yakA-mediated developmental signaling pathway, and expression of the extracellular matrix/cell adhesion proteins (DdCAD and csA). This resulted in delayed chemotaxis, adhesion, and development of the organism. In contrast to the inhibitory effects on developmental genes, curcumin induced gstA gene expression, overall GST activity, and generated production of reactive oxygen species. These studies expand our knowledge of developmental and biochemical signaling influenced by curcumin, and lends greater consideration of GST enzyme function in eukaryotic cell signaling, development, and differentiation.

Curcumin inhibits development and cell adhesion in Dictyostelium discoideum: Implications for YakA signaling and GST enzyme function

International Nuclear Information System (INIS)

Garige, Mamatha; Walters, Eric

2015-01-01

The molecular basis for nutraceutical properties of the polyphenol curcumin (Curcuma longa, Turmeric) is complex, affecting multiple factors that regulate cell signaling and homeostasis. Here, we report the effect of curcumin on cellular and developmental mechanisms in the eukaryotic model, Dictyostelium discoideum. Dictyostelium proliferation was inhibited in the presence of curcumin, which also suppressed the prestarvation marker, discoidin I, members of the yakA-mediated developmental signaling pathway, and expression of the extracellular matrix/cell adhesion proteins (DdCAD and csA). This resulted in delayed chemotaxis, adhesion, and development of the organism. In contrast to the inhibitory effects on developmental genes, curcumin induced gstA gene expression, overall GST activity, and generated production of reactive oxygen species. These studies expand our knowledge of developmental and biochemical signaling influenced by curcumin, and lends greater consideration of GST enzyme function in eukaryotic cell signaling, development, and differentiation.

The actinome of Dictyostelium discoideum in comparison to actins and actin-related proteins from other organisms.

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Jayabalan M Joseph

Full Text Available Actin belongs to the most abundant proteins in eukaryotic cells which harbor usually many conventional actin isoforms as well as actin-related proteins (Arps. To get an overview over the sometimes confusing multitude of actins and Arps, we analyzed the Dictyostelium discoideum actinome in detail and compared it with the genomes from other model organisms. The D. discoideum actinome comprises 41 actins and actin-related proteins. The genome contains 17 actin genes which most likely arose from consecutive gene duplications, are all active, in some cases developmentally regulated and coding for identical proteins (Act8-group. According to published data, the actin fraction in a D. discoideum cell consists of more than 95% of these Act8-type proteins. The other 16 actin isoforms contain a conventional actin motif profile as well but differ in their protein sequences. Seven actin genes are potential pseudogenes. A homology search of the human genome using the most typical D. discoideum actin (Act8 as query sequence finds the major actin isoforms such as cytoplasmic beta-actin as best hit. This suggests that the Act8-group represents a nearly perfect actin throughout evolution. Interestingly, limited data from D. fasciculatum, a more ancient member among the social amoebae, show different relationships between conventional actins. The Act8-type isoform is most conserved throughout evolution. Modeling of the putative structures suggests that the majority of the actin-related proteins is functionally unrelated to canonical actin. The data suggest that the other actin variants are not necessary for the cytoskeleton itself but rather regulators of its dynamical features or subunits in larger protein complexes.

Uncovering a Role for the Tail of the Dictyostelium discoideum SadA Protein in Cell-Substrate Adhesion ▿ †

OpenAIRE

Kowal, Anthony S.; Chisholm, Rex L.

2011-01-01

Previous work from our laboratory showed that the Dictyostelium discoideum SadA protein plays a central role in cell-substrate adhesion. SadA null cells exhibit a loss of adhesion, a disrupted actin cytoskeleton, and a cytokinesis defect. How SadA mediates these phenotypes is unknown. This work addresses the mechanism of SadA function, demonstrating an important role for the C-terminal cytoplasmic tail in SadA function. We found that a SadA tailless mutant was unable to rescue the sadA adhesi...

Cytochemical study of the nucleolus of the slime mold Dictyostelium discoideum

International Nuclear Information System (INIS)

Benichou, J.C.; Quiviger, B.; Ryter, A.

1983-01-01

The nucleus of the slime mold Dictyostelium discoideum is characterized by the presence of several large dense masses which are all in tight contact with the nuclear membrane. These dense masses, considered as nucleoli, present a rather homogeneous texture, in which dense chromatin, fibrillar, and granular material are not easily detected. The autoradiographic study of [ 3 H]uridine pulse-labeled cells showed that the majority of the silver grains were located inside these masses. The use of EDTA regressive-staining, acetylation and enzymatic digestion indicated that they are mostly composed of RNP and are totally devoid of dense chromatin as the rest of the nucleus is. After treatment with actinomycin D, fibrillar and granular material segregated but no chromatin could be found. All these observations confirmed that the dense masses correspond to nucleoli despite their peculiar ultrastructure. It can also be concluded that this type of nucleoli cannot be considered as a taxonomic character of the slime molds because it does not exist in all slime molds and was observed in some dinoflagellates, and ascomycetes

Clues to γ-secretase, huntingtin and Hirano body normal function using the model organism Dictyostelium discoideum

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Myre Michael A

2012-04-01

Full Text Available Abstract Many neurodegenerative disorders, although related by their destruction of brain function, display remarkable cellular and/or regional pathogenic specificity likely due to a deregulated functionality of the mutant protein. However, neurodegenerative disease genes, for example huntingtin (HTT, the ataxins, the presenilins (PSEN1/PSEN2 are not simply localized to neurons but are ubiquitously expressed throughout peripheral tissues; it is therefore paramount to properly understand the earliest precipitating events leading to neuronal pathogenesis to develop effective long-term therapies. This means, in no unequivocal terms, it is crucial to understand the gene's normal function. Unfortunately, many genes are often essential for embryogenesis which precludes their study in whole organisms. This is true for HTT, the β-amyloid precursor protein (APP and presenilins, responsible for early onset Alzheimer's disease (AD. To better understand neurological disease in humans, many lower and higher eukaryotic models have been established. So the question arises: how reasonable is the use of organisms to study neurological disorders when the model of choice does not contain neurons? Here we will review the surprising, and novel emerging use of the model organism Dictyostelium discoideum, a species of soil-living amoeba, as a valuable biomedical tool to study the normal function of neurodegenerative genes. Historically, the evidence on the usefulness of simple organisms to understand the etiology of cellular pathology cannot be denied. But using an organism without a central nervous system to understand diseases of the brain? We will first introduce the life cycle of Dictyostelium, the presence of many disease genes in the genome and how it has provided unique opportunities to identify mechanisms of disease involving actin pathologies, mitochondrial disease, human lysosomal and trafficking disorders and host-pathogen interactions. Secondly, I will

Assets of the non-pathogenic microorganism Dictyostelium discoideum as a model for the study of eukaryotic extracellular vesicles [v1; ref status: indexed, http://f1000r.es/pa

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Irène Tatischeff

2013-03-01

Full Text Available Dictyostelium discoideum microvesicles have recently been presented as a valuable model for eukaryotic extracellular vesicles. Here, the advantages of D. discoideum for unraveling important biological functions of extracellular vesicles in general are detailed. D. discoideum, a non-pathogenic eukaryotic microorganism, belongs to a billion-year-old Amoeboza lineage, which diverged from the animal-fungal lineage after the plant animal-split. During growth and early starvation-induced development, it presents analogies with lymphocytes and macrophages with regard to motility and phagocytosis capability, respectively. Its 6-chromosome genome codes for about 12,500 genes, some showing analogies with human genes. The presence of extracellular vesicles during cell growth has been evidenced as a detoxification mechanism of various structurally unrelated drugs. Controls led to the discovery of constitutive extracellular vesicle secretion in this microorganism, which was an important point. It means that the secretion of extracellular vesicles occurs, in the absence of any drug, during both cell growth and early development. This constitutive secretion of D. discoideum cells is very likely to play a role in intercellular communication. The detoxifying secreted vesicles, which can transport drugs outside the cells, can also act as "Trojan horses", capable of transferring these drugs not only into naïve D. discoideum cells, but into human cells as well. Therefore, these extracellular vesicles were proposed as a new biological drug delivery tool. Moreover, Dictyostelium, chosen by the NIH (USA as a new model organism for biomedical research, has already been used for studying some human diseases. These cells, which are much easier to manipulate than human cells, can be easily designed in simple conditioned medium experiments. Owing to the increasing consensus that extracellular vesicles are probably important mediators of intercellular communication, D. discoideum

Loss of Cln3 Function in the Social Amoeba Dictyostelium discoideum Causes Pleiotropic Effects That Are Rescued by Human CLN3

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Huber, Robert J.

2014-01-01

The neuronal ceroid lipofuscinoses (NCL) are a group of inherited, severe neurodegenerative disorders also known as Batten disease. Juvenile NCL (JNCL) is caused by recessive loss-of-function mutations in CLN3, which encodes a transmembrane protein that regulates endocytic pathway trafficking, though its primary function is not yet known. The social amoeba Dictyostelium discoideum is increasingly utilized for neurological disease research and is particularly suited for investigation of protein function in trafficking. Therefore, here we establish new overexpression and knockout Dictyostelium cell lines for JNCL research. Dictyostelium Cln3 fused to GFP localized to the contractile vacuole system and to compartments of the endocytic pathway. cln3− cells displayed increased rates of proliferation and an associated reduction in the extracellular levels and cleavage of the autocrine proliferation repressor, AprA. Mid- and late development of cln3− cells was precocious and cln3− slugs displayed increased migration. Expression of either Dictyostelium Cln3 or human CLN3 in cln3− cells suppressed the precocious development and aberrant slug migration, which were also suppressed by calcium chelation. Taken together, our results show that Cln3 is a pleiotropic protein that negatively regulates proliferation and development in Dictyostelium. This new model system, which allows for the study of Cln3 function in both single cells and a multicellular organism, together with the observation that expression of human CLN3 restores abnormalities in Dictyostelium cln3− cells, strongly supports the use of this new model for JNCL research. PMID:25330233

Loss of Cln3 function in the social amoeba Dictyostelium discoideum causes pleiotropic effects that are rescued by human CLN3.

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Robert J Huber

Full Text Available The neuronal ceroid lipofuscinoses (NCL are a group of inherited, severe neurodegenerative disorders also known as Batten disease. Juvenile NCL (JNCL is caused by recessive loss-of-function mutations in CLN3, which encodes a transmembrane protein that regulates endocytic pathway trafficking, though its primary function is not yet known. The social amoeba Dictyostelium discoideum is increasingly utilized for neurological disease research and is particularly suited for investigation of protein function in trafficking. Therefore, here we establish new overexpression and knockout Dictyostelium cell lines for JNCL research. Dictyostelium Cln3 fused to GFP localized to the contractile vacuole system and to compartments of the endocytic pathway. cln3- cells displayed increased rates of proliferation and an associated reduction in the extracellular levels and cleavage of the autocrine proliferation repressor, AprA. Mid- and late development of cln3- cells was precocious and cln3- slugs displayed increased migration. Expression of either Dictyostelium Cln3 or human CLN3 in cln3- cells suppressed the precocious development and aberrant slug migration, which were also suppressed by calcium chelation. Taken together, our results show that Cln3 is a pleiotropic protein that negatively regulates proliferation and development in Dictyostelium. This new model system, which allows for the study of Cln3 function in both single cells and a multicellular organism, together with the observation that expression of human CLN3 restores abnormalities in Dictyostelium cln3- cells, strongly supports the use of this new model for JNCL research.

eIF2α Kinases Control Chalone Production in Dictyostelium discoideum ▿

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Bowman, Robert L.; Xiong, Yanhua; Kirsten, Janet H.; Singleton, Charles K.

2011-01-01

Growing Dictyostelium cells secrete CfaD and AprA, two proteins that have been characterized as chalones. They exist within a high-molecular-weight complex that reversibly inhibits cell proliferation, but not growth, via cell surface receptors and a signaling pathway that includes G proteins. How the production of these two proteins is regulated is unknown. Dictyostelium cells possess three GCN2-type eukaryotic initiation factor 2 α subunit (eIF2α) kinases, proteins that phosphorylate the translational initiation factor eIF2α and possess a tRNA binding domain involved in their regulation. The Dictyostelium kinases have been shown to function during development in regulating several processes. We show here that expression of an unregulated, activated kinase domain greatly inhibits cell proliferation. The inhibitory effect on proliferation is not due to a general inhibition of translation. Instead, it is due to enhanced production of a secreted factor(s). Indeed, extracellular CfaD and AprA proteins, but not their mRNAs, are overproduced in cells expressing the activated kinase domain. The inhibition of proliferation is not seen when the activated kinase domain is expressed in cells lacking CfaD or AprA or in cells that contain a nonphosphorylatable eIF2α. We conclude that production of the chalones CfaD and AprA is translationally regulated by eIF2α phosphorylation. Both proteins are upregulated at the culmination of development, and this enhanced production is lacking in a strain that possesses a nonphosphorylatable eIF2α. PMID:21278229

RNA synthesis during germination of UV-irradiated Dictyostelium discoideum spores

International Nuclear Information System (INIS)

Okaichi, Kumio

1987-01-01

UV irradiation to the spores of Dictyostelium discoideum NC4 resulted in a more prolonged delay of amoeba-emergence from swollen spores with increasing UV fluence. During the germination, an inhibition of total RNA synthesis and a shift of stage of maximum RNA synthesis to the later period were observed. The maximum poly(A) + RNA synthetic activity was found on an early stage of amoeba-emergence prior about 1 h to the beginning of rRNA synthesis in unirradiated spore germination; but, in UV-irradiated spore germination, the stage of maximum poly(A) + RNA synthesis shifted to the later stage of germination with increasing UV fluence. A decreased synthesis of poly(A) + RNA and a severe inhibition of rRNA synthesis were observed on UV-irradiated and germinated spores, but no significant inhibition of 4 - 5 S RNA synthesis was detected. Actinomycin D suppressed almost completely the rRNA synthesis of emerged amoebae but the drug apparently did not affect the emergence of amoebae at any stage of germination. It was postulated that the delay of amoeba-emergence in UV-irradiated spore must be mainly due to the shift of the stage of maximum synthesis of poly(A) + RNA to the later stage of germination. (author)

Arachidonic acid is a chemoattractant for Dictyostelium discoideum ...

Indian Academy of Sciences (India)

SEARCHU

binding proteins and calmodulin-dependent phosphorylation linked to calmodulin-dependent chemotaxis to folic and cAMP in Dictyostelium; Cell Signal 13 575–584. Gerisch G and Hess B 1974 Cyclic-AMP-controlled oscillations in suspended Dictyostelium cells: Their relation to morphogenetic cell interactions; Proc. Natl.

Characterization of a 1,4-{beta}-D-glucan synthase from Dictyostelium discoideum. Progress report, May 1990--January 1992

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Blanton, R.L.

1992-01-15

Various aspects of research concerning Dictyostelium discoideum are presented. The initial focus of this project was upon: the characterization of potential probes for the cellulose synthase (antibody and nucleic acid), the determination of the cultural induction conditions of cellulose synthesis, the solubilization of the enzyme activity, the development of a non-inhibitory disruption buffer, the generation and isolation of mutant strains deficient in cellulose synthesis, and the development of the capability to determine the degree of polymerization of the in vitro product. I have briefly summarized our most significant findings with only selected data sets being shown in this report in the interest of brevity.

Overexpression of TOR (target of rapamycin) inhibits cell proliferation in Dictyostelium discoideum.

Science.gov (United States)

Swer, Pynskhem Bok; Mishra, Himanshu; Lohia, Rakhee; Saran, Shweta

2016-05-01

TOR (target of rapamycin) protein kinase acts as a central controller of cell growth and development of an organism. Present study was undertaken to find the expression pattern and role of TOR during growth and development of Dictyostelium discoideum. Failures to generate either knockout and/or knockdown mutants indicate that interference with its levels led to cellular defects. Thus, the effects of TOR (DDB_G0281569) overexpression specifically, cells expressing Dd(Δ211-TOR)-Eyfp mutant was analyzed. Elevated expression of (Δ211-TOR)-Eyfp reduced both cell size and cell proliferation. DdTOR was found to be closer to fungus. mRNA level of TOR was found maximally in the freshly starved/aggregate cells that gradually declined. This was also strengthened by the expression patterns observed by in situ and the analysis of β-galactosidase reporter driven by the putative TOR promoter. The TOR protein was found to be highest at the aggregate stage. The fusion protein, (Δ211-TOR)-Eyfp was localized to the cell membrane, cytosol, and the nucleus. We suggest, DdTOR to be an essential protein and high TOR expression inhibits cell proliferation. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Evaluating Different Virulence Traits of Klebsiella pneumoniae Using Dictyostelium discoideum and Zebrafish Larvae as Host Models

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Andrés E. Marcoleta

2018-02-01

Full Text Available Multiresistant and invasive hypervirulent Klebsiella pneumoniae strains have become one of the most urgent bacterial pathogen threats. Recent analyses revealed a high genomic plasticity of this species, harboring a variety of mobile genetic elements associated with virulent strains, encoding proteins of unknown function whose possible role in pathogenesis have not been addressed. K. pneumoniae virulence has been studied mainly in animal models such as mice and pigs, however, practical, financial, ethical and methodological issues limit the use of mammal hosts. Consequently, the development of simple and cost-effective experimental approaches with alternative host models is needed. In this work we described the use of both, the social amoeba and professional phagocyte Dictyostelium discoideum and the fish Danio rerio (zebrafish as surrogate host models to study K. pneumoniae virulence. We compared three K. pneumoniae clinical isolates evaluating their resistance to phagocytosis, intracellular survival, lethality, intestinal colonization, and innate immune cells recruitment. Optical transparency of both host models permitted studying the infective process in vivo, following the Klebsiella-host interactions through live-cell imaging. We demonstrated that K. pneumoniae RYC492, but not the multiresistant strains 700603 and BAA-1705, is virulent to both host models and elicits a strong immune response. Moreover, this strain showed a high resistance to phagocytosis by D. discoideum, an increased ability to form biofilms and a more prominent and irregular capsule. Besides, the strain 700603 showed the unique ability to replicate inside amoeba cells. Genomic comparison of the K. pneumoniae strains showed that the RYC492 strain has a higher overall content of virulence factors although no specific genes could be linked to its phagocytosis resistance, nor to the intracellular survival observed for the 700603 strain. Our results indicate that both zebrafish

Excitable signal relay in Dictyostelium discoideum

Science.gov (United States)

Mestler, Troy; Schwab, David; Mehta, Pankaj; Gregor, Thomas

2011-03-01

The social amoeba D. discoideum transitions when starved from a collection of individual cells into a multicellular spore-complex. During this process, amoebae display several interesting phenomena including intercellular signaling, pattern formation, and cell differentiation. At the heart of these phenomena is the exchange of the signaling molecule cyclic-AMP, which has previously been extensively studied using a variety of indirect methods. Here we employ a sensor that uses a compound fluorescent protein whose emission spectrum changes in the presence of bound cyclic AMP to directly monitor, in real time and in vivo, intracellular cAMP concentrations. We use cells expressing this sensor in microchemostats to study intracellular cAMP concentrations at the single-cell level in response to precise, dynamically-controlled external cAMP stimulation. Specifically, we show that these cells display excitability much like that found in neurons and agree experimentally quite well with a modified FitzHugh-Nagumo dynamical systems model. This single-cell model sets groundwork for a comprehensive multicellular model that promises to explain emergent behavior in D. discoideum.

1H, 15N and 13C assignments of domain 5 of Dictyostelium discoideum gelation factor (ABP-120) in its native and 8M urea-denatured states.

Science.gov (United States)

Hsu, Shang-Te Danny; Cabrita, Lisa D; Christodoulou, John; Dobson, Christopher M

2009-06-01

The gelation factor from Dictyostelium discoideum (ABP-120) is an actin binding protein consisting of six immunoglobulin (Ig) domains in the C-terminal rod domain. We have recently used the pair of domains 5 and 6 of ABP-120 as a model system for studying multi-domain nascent chain folding on the ribosome. Here we present the NMR assignments of domain 5 in its native and 8M urea-denatured states.

Dicty_cDB: [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VF (Link to library) VFB606 (Link to dictyBase) - - - Contig-U16382-1 VFB606E (Link...) Clone ID VFB606 (Link to dictyBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U16382-1 Ori...ucing significant alignments: (bits) Value N X03281 |X03281.1 Dictyostelium discoideum gene for actin A8. 22...28 0.0 1 AC116957 |AC116957.2 Dictyostelium discoideum chromosome 2 map 1685067-2...2028 0.0 1 AC115579 |AC115579.2 Dictyostelium discoideum chromosome 2 map 4915084-5005461 strain AX4, comple

Dictyostelium discoideum: mutants in the biosynthesis of the lipid-linked precursor of N-linked oligosaccharides

International Nuclear Information System (INIS)

Freeze, H.; Willies, L.; Hamilton, S.

1986-01-01

The lysosomal enzymes of Dictyostelium discoideum share highly immunogenic oligosaccharides which contain multiple Man-6-SO 4 residues. Two mutant strains which lack the shared antigenic determinant were analyzed in an attempt to identify the primary defect in each. [ 3 H]Man labelled N-linked oligosaccharides of secreted glycoproteins were released by Endo/PNGaseF digestion and analyzed. Both of the mutant strains produced smaller, less sulfated oligosaccharides compared to the wild-type, yet both still contained considerable amounts of Man-6-SO 4 . The size of the precursor lipid-linked oligosaccharide from the wild-type is consistent with a Glc 3 Man 9 GlcNAc 2 structure, while those from both of the mutants have an oligosaccharide the size of Man 5 GlcNAc 2 . The authors conclude that both of the mutants are defective in the biosynthesis of the precursor oligosaccharide. Both oligosaccharides from the mutants contain a tri-mannosyl core and are not glucosylated. Two of the five Man residues are released by a 1,2 specific α mannosidase. Based on the size and mannosidase digestions the authors conclude that 4/5 of the Man residues on the α1,6 branch of the β-linked Man residues are missing. Thus, these residues must be required to define the shared antigenic determinant

Fluorographic detection of tritiated glycopeptides and oligosaccharides separated on polyacrylamide gels: analysis of glycans from Dictyostelium discoideum glycoproteins

International Nuclear Information System (INIS)

Prem Das, O.; Henderson, E.J.

1986-01-01

Previous workers have shown that oligosaccharides and glycopeptides can be separated by electrophoresis in buffers containing borate ions. However, normal fluorography of tritium-labeled structures cannot be performed because the glycans are soluble and can diffuse during equilibration with scintillants. This problem has been circumvented by equilibration of the gel with 2,5-diphenyloxazole (PPO) prior to electrophoresis. The presence of PPO in the gel during electrophoresis does not alter mobility of the glycopeptides and oligosaccharides. After electrophoresis, the gel is simply dried and fluorography performed. This allows sensitive and precise comparisons of labeled samples in parallel lanes of a slab gel and, since mobilities are highly reproducible, between different gels. The procedure is preparative in that after fluorography the gel bands can be quantitatively eluted for further study, without any apparent modification by the procedure. In this report, the procedure is illustrated by fractionation of both neutral and anionic glycopeptides produced by the cellular slime mold Dictyostelium discoideum

Dicty_cDB: [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VF (Link to library) VFB645 (Link to dictyBase) - - - Contig-U16382-1 VFB645P (Link... to Original site) VFB645F 619 VFB645Z 413 VFB645P 1032 - - Show VFB645 Library VF (Link to library) Clone ID VFB645 (Link to dict...yBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U16382-1 Original site URL http://dict...mology vs DNA Score E Sequences producing significant alignments: (bits) Value N X03281 |X03281.1 Dictyostel...ium discoideum gene for actin A8. 1174 0.0 2 AC116957 |AC116957.2 Dictyostelium discoideum chromosome 2 map

Dicty_cDB: [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VF (Link to library) VFB741 (Link to dictyBase) - - - Contig-U16382-1 VFB741P (Link... to Original site) VFB741F 545 VFB741Z 675 VFB741P 1220 - - Show VFB741 Library VF (Link to library) Clone ID VFB741 (Link to dict...yBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U16382-1 Original site URL http://dict...bits) Value N X03281 |X03281.1 Dictyostelium discoideum gene for actin A8. 1166 0....0 2 AC116957 |AC116957.2 Dictyostelium discoideum chromosome 2 map 1685067-2090751 strain AX4, complete seq

Dicty_cDB: [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VF (Link to library) VFB710 (Link to dictyBase) - - - Contig-U16382-1 VFB710P (Link... to Original site) VFB710F 123 VFB710Z 689 VFB710P 812 - - Show VFB710 Library VF (Link to library) Clone ID VFB710 (Link to dict...yBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U16382-1 Original site URL http://dict... significant alignments: (bits) Value N X03281 |X03281.1 Dictyostelium discoideum... gene for actin A8. 1308 0.0 2 AC116957 |AC116957.2 Dictyostelium discoideum chromosome 2 map 1685067-209075

dictyExpress: a Dictyostelium discoideum gene expression database with an explorative data analysis web-based interface

Science.gov (United States)

Rot, Gregor; Parikh, Anup; Curk, Tomaz; Kuspa, Adam; Shaulsky, Gad; Zupan, Blaz

2009-01-01

Background Bioinformatics often leverages on recent advancements in computer science to support biologists in their scientific discovery process. Such efforts include the development of easy-to-use web interfaces to biomedical databases. Recent advancements in interactive web technologies require us to rethink the standard submit-and-wait paradigm, and craft bioinformatics web applications that share analytical and interactive power with their desktop relatives, while retaining simplicity and availability. Results We have developed dictyExpress, a web application that features a graphical, highly interactive explorative interface to our database that consists of more than 1000 Dictyostelium discoideum gene expression experiments. In dictyExpress, the user can select experiments and genes, perform gene clustering, view gene expression profiles across time, view gene co-expression networks, perform analyses of Gene Ontology term enrichment, and simultaneously display expression profiles for a selected gene in various experiments. Most importantly, these tasks are achieved through web applications whose components are seamlessly interlinked and immediately respond to events triggered by the user, thus providing a powerful explorative data analysis environment. Conclusion dictyExpress is a precursor for a new generation of web-based bioinformatics applications with simple but powerful interactive interfaces that resemble that of the modern desktop. While dictyExpress serves mainly the Dictyostelium research community, it is relatively easy to adapt it to other datasets. We propose that the design ideas behind dictyExpress will influence the development of similar applications for other model organisms. PMID:19706156

The p21-activated kinase (PAK family member PakD is required for chemorepulsion and proliferation inhibition by autocrine signals in Dictyostelium discoideum.

Directory of Open Access Journals (Sweden)

Jonathan E Phillips

Full Text Available In Dictyostelium discoideum, the secreted proteins AprA and CfaD function as reporters of cell density and regulate cell number by inhibiting proliferation at high cell densities. AprA also functions to disperse groups of cells at high density by acting as a chemorepellent. However, the signal transduction pathways associated with AprA and CfaD are not clear, and little is known about how AprA affects the cytoskeleton to regulate cell movement. We found that the p21-activated kinase (PAK family member PakD is required for both the proliferation-inhibiting activity of AprA and CfaD and the chemorepellent activity of AprA. Similar to cells lacking AprA or CfaD, cells lacking PakD proliferate to a higher cell density than wild-type cells. Recombinant AprA and CfaD inhibit the proliferation of wild-type cells but not cells lacking PakD. Like AprA and CfaD, PakD affects proliferation but does not significantly affect growth (the accumulation of mass on a per-nucleus basis. In contrast to wild-type cells, cells lacking PakD are not repelled from a source of AprA, and colonies of cells lacking PakD expand at a slower rate than wild-type cells, indicating that PakD is required for AprA-mediated chemorepulsion. A PakD-GFP fusion protein localizes to an intracellular punctum that is not the nucleus or centrosome, and PakD-GFP is also occasionally observed at the rear cortex of moving cells. Vegetative cells lacking PakD show excessive actin-based filopodia-like structures, suggesting that PakD affects actin dynamics, consistent with previously characterized roles of PAK proteins in actin regulation. Together, our results implicate PakD in AprA/CfaD signaling and show that a PAK protein is required for proper chemorepulsive cell movement in Dictyostelium.

The p21-activated kinase (PAK) family member PakD is required for chemorepulsion and proliferation inhibition by autocrine signals in Dictyostelium discoideum.

Science.gov (United States)

Phillips, Jonathan E; Gomer, Richard H

2014-01-01

In Dictyostelium discoideum, the secreted proteins AprA and CfaD function as reporters of cell density and regulate cell number by inhibiting proliferation at high cell densities. AprA also functions to disperse groups of cells at high density by acting as a chemorepellent. However, the signal transduction pathways associated with AprA and CfaD are not clear, and little is known about how AprA affects the cytoskeleton to regulate cell movement. We found that the p21-activated kinase (PAK) family member PakD is required for both the proliferation-inhibiting activity of AprA and CfaD and the chemorepellent activity of AprA. Similar to cells lacking AprA or CfaD, cells lacking PakD proliferate to a higher cell density than wild-type cells. Recombinant AprA and CfaD inhibit the proliferation of wild-type cells but not cells lacking PakD. Like AprA and CfaD, PakD affects proliferation but does not significantly affect growth (the accumulation of mass) on a per-nucleus basis. In contrast to wild-type cells, cells lacking PakD are not repelled from a source of AprA, and colonies of cells lacking PakD expand at a slower rate than wild-type cells, indicating that PakD is required for AprA-mediated chemorepulsion. A PakD-GFP fusion protein localizes to an intracellular punctum that is not the nucleus or centrosome, and PakD-GFP is also occasionally observed at the rear cortex of moving cells. Vegetative cells lacking PakD show excessive actin-based filopodia-like structures, suggesting that PakD affects actin dynamics, consistent with previously characterized roles of PAK proteins in actin regulation. Together, our results implicate PakD in AprA/CfaD signaling and show that a PAK protein is required for proper chemorepulsive cell movement in Dictyostelium.

Quadruplexes in 'Dicty': crystal structure of a four-quartet G-quadruplex formed by G-rich motif found in the Dictyostelium discoideum genome.

Science.gov (United States)

Guédin, Aurore; Lin, Linda Yingqi; Armane, Samir; Lacroix, Laurent; Mergny, Jean-Louis; Thore, Stéphane; Yatsunyk, Liliya A

2018-06-01

Guanine-rich DNA has the potential to fold into non-canonical G-quadruplex (G4) structures. Analysis of the genome of the social amoeba Dictyostelium discoideum indicates a low number of sequences with G4-forming potential (249-1055). Therefore, D. discoideum is a perfect model organism to investigate the relationship between the presence of G4s and their biological functions. As a first step in this investigation, we crystallized the dGGGGGAGGGGTACAGGGGTACAGGGG sequence from the putative promoter region of two divergent genes in D. discoideum. According to the crystal structure, this sequence folds into a four-quartet intramolecular antiparallel G4 with two lateral and one diagonal loops. The G-quadruplex core is further stabilized by a G-C Watson-Crick base pair and a A-T-A triad and displays high thermal stability (Tm > 90°C at 100 mM KCl). Biophysical characterization of the native sequence and loop mutants suggests that the DNA adopts the same structure in solution and in crystalline form, and that loop interactions are important for the G4 stability but not for its folding. Four-tetrad G4 structures are sparse. Thus, our work advances understanding of the structural diversity of G-quadruplexes and yields coordinates for in silico drug screening programs and G4 predictive tools.

Dicty_cDB: [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VF (Link to library) VFB770 (Link to dictyBase) - - - Contig-U16382-1 VFB770E (Link...) Clone ID VFB770 (Link to dictyBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U16382-1 Ori...logy vs DNA Score E Sequences producing significant alignments: (bits) Value N X03281 |X03281.1 Dictyosteliu...m discoideum gene for actin A8. 2230 0.0 1 AC116957 |AC116957.2 Dictyostelium discoideum chromosome 2 map 16...deum actin 15 gene, complete cds. 2030 0.0 1 AC115579 |AC115579.2 Dictyostelium discoideum chromosome 2 map

The TOM Complex of Amoebozoans: the Cases of the Amoeba Acanthamoeba castellanii and the Slime Mold Dictyostelium discoideum.

Science.gov (United States)

Wojtkowska, Małgorzata; Buczek, Dorota; Stobienia, Olgierd; Karachitos, Andonis; Antoniewicz, Monika; Slocinska, Małgorzata; Makałowski, Wojciech; Kmita, Hanna

2015-07-01

Protein import into mitochondria requires a wide variety of proteins, forming complexes in both mitochondrial membranes. The TOM complex (translocase of the outer membrane) is responsible for decoding of targeting signals, translocation of imported proteins across or into the outer membrane, and their subsequent sorting. Thus the TOM complex is regarded as the main gate into mitochondria for imported proteins. Available data indicate that mitochondria of representative organisms from across the major phylogenetic lineages of eukaryotes differ in subunit organization of the TOM complex. The subunit organization of the TOM complex in the Amoebozoa is still elusive, so we decided to investigate its organization in the soil amoeba Acanthamoeba castellanii and the slime mold Dictyostelium discoideum. They represent two major subclades of the Amoebozoa: the Lobosa and Conosa, respectively. Our results confirm the presence of Tom70, Tom40 and Tom7 in the A. castellanii and D. discoideum TOM complex, while the presence of Tom22 and Tom20 is less supported. Interestingly, the Tom proteins display the highest similarity to Opisthokonta cognate proteins, with the exception of Tom40. Thus representatives of two major subclades of the Amoebozoa appear to be similar in organization of the TOM complex, despite differences in their lifestyle. Copyright © 2015 The Authors. Published by Elsevier GmbH.. All rights reserved.

An Autocrine Proliferation Repressor Regulates Dictyostelium discoideum Proliferation and Chemorepulsion Using the G Protein-Coupled Receptor GrlH.

Science.gov (United States)

Tang, Yu; Wu, Yuantai; Herlihy, Sarah E; Brito-Aleman, Francisco J; Ting, Jose H; Janetopoulos, Chris; Gomer, Richard H

2018-02-13

In eukaryotic microbes, little is known about signals that inhibit the proliferation of the cells that secrete the signal, and little is known about signals (chemorepellents) that cause cells to move away from the source of the signal. Autocrine proliferation repressor protein A (AprA) is a protein secreted by the eukaryotic microbe Dictyostelium discoideum AprA is a chemorepellent for and inhibits the proliferation of D. discoideum We previously found that cells sense AprA using G proteins, suggesting the existence of a G protein-coupled AprA receptor. To identify the AprA receptor, we screened mutants lacking putative G protein-coupled receptors. We found that, compared to the wild-type strain, cells lacking putative receptor GrlH ( grlH¯ cells) show rapid proliferation, do not have large numbers of cells moving away from the edges of colonies, are insensitive to AprA-induced proliferation inhibition and chemorepulsion, and have decreased AprA binding. Expression of GrlH in grlH¯ cells ( grlH¯/grlH OE ) rescues the phenotypes described above. These data indicate that AprA signaling may be mediated by GrlH in D. discoideum IMPORTANCE Little is known about how eukaryotic cells can count themselves and thus regulate the size of a tissue or density of cells. In addition, little is known about how eukaryotic cells can sense a repellant signal and move away from the source of the repellant, for instance, to organize the movement of cells in a developing embryo or to move immune cells out of a tissue. In this study, we found that a eukaryotic microbe uses G protein-coupled receptors to mediate both cell density sensing and chemorepulsion. Copyright © 2018 Tang et al.

Inhibitors of Mycobacterium marinum virulence identified in a Dictyostelium discoideum host model.

Directory of Open Access Journals (Sweden)

Hajer Ouertatani-Sakouhi

Full Text Available Tuberculosis remains one of the major threats to public health worldwide. Given the prevalence of multi drug resistance (MDR in Mycobacterium tuberculosis strains, there is a strong need to develop new anti-mycobacterial drugs with modes of action distinct from classical antibiotics. Inhibitors of mycobacterial virulence might target new molecular processes and may represent a potential new therapeutic alternative. In this study, we used a Dictyostelium discoideum host model to assess virulence of Mycobacterium marinum and to identify compounds inhibiting mycobacterial virulence. Among 9995 chemical compounds, we selected 12 inhibitors of mycobacterial virulence that do not inhibit mycobacterial growth in synthetic medium. Further analyses revealed that 8 of them perturbed functions requiring an intact mycobacterial cell wall such as sliding motility, bacterial aggregation or cell wall permeability. Chemical analogs of two compounds were analyzed. Chemical modifications altered concomitantly their effect on sliding motility and on mycobacterial virulence, suggesting that the alteration of the mycobacterial cell wall caused the loss of virulence. We characterized further one of the selected compounds and found that it inhibited the ability of mycobacteria to replicate in infected cells. Together these results identify new antimycobacterial compounds that represent new tools to unravel the molecular mechanisms controlling mycobacterial pathogenicity. The isolation of compounds with anti-virulence activity is the first step towards developing new antibacterial treatments.

The human homologue of Dictyostelium discoideum phg1A is expressed by human metastatic melanoma cells.

Science.gov (United States)

Lozupone, Francesco; Perdicchio, Maurizio; Brambilla, Daria; Borghi, Martina; Meschini, Stefania; Barca, Stefano; Marino, Maria Lucia; Logozzi, Mariantonia; Federici, Cristina; Iessi, Elisabetta; de Milito, Angelo; Fais, Stefano

2009-12-01

Tumour cannibalism is a characteristic of malignancy and metastatic behaviour. This atypical phagocytic activity is a crucial survival option for tumours in conditions of low nutrient supply, and has some similarities to the phagocytic activity of unicellular microorganisms. In fact, Dictyostelium discoideum has been used widely as a model to study phagocytosis. Recently, phg1A has been described as a protein that is primarily involved in the phagocytic process of this microorganism. The closest human homologue to phg1A is transmembrane 9 superfamily protein member 4 (TM9SF4). Here, we report that TM9SF4 is highly expressed in human malignant melanoma cells deriving from metastatic lesions, whereas it is undetectable in healthy human tissues and cells. TM9SF4 is predominantly expressed in acidic vesicles of melanoma cells, in which it co-localizes with the early endosome antigens Rab5 and early endosome antigen 1. TM9SF4 silencing induced marked inhibition of cannibal activity, which is consistent with a derangement of intracellular pH gradients, with alkalinization of acidic vesicles and acidification of the cell cytosol. We propose TM9SF4 as a new marker of malignancy, representing a potential new target for anti-tumour strategies with a specific role in tumour cannibalism and in the establishment of a metastatic phenotype.

Role of phospholipase C in Dictyostelium : Formation of inositol 1,4,5-trisphosphate and normal development in cells lacking phospholipase C activity

NARCIS (Netherlands)

Drayer, A. Lyndsay; Kaay, Jeroen van der; Mayr, Georg W.; Haastert, Peter J.M. van

1994-01-01

The micro-organism Dictyostelium uses extracellular cAMP to induce chemotaxis and cell differentiation. Signals are transduced via surface receptors, which activate G proteins, to effector enzymes. The deduced protein sequence of Dictyostelium discoideum phosphabidylinositol-specific phospholipase C

MicroRNAs in Amoebozoa: deep sequencing of the small RNA population in the social amoeba Dictyostelium discoideum reveals developmentally regulated microRNAs.

Science.gov (United States)

Avesson, Lotta; Reimegård, Johan; Wagner, E Gerhart H; Söderbom, Fredrik

2012-10-01

The RNA interference machinery has served as a guardian of eukaryotic genomes since the divergence from prokaryotes. Although the basic components have a shared origin, silencing pathways directed by small RNAs have evolved in diverse directions in different eukaryotic lineages. Micro (mi)RNAs regulate protein-coding genes and play vital roles in plants and animals, but less is known about their functions in other organisms. Here, we report, for the first time, deep sequencing of small RNAs from the social amoeba Dictyostelium discoideum. RNA from growing single-cell amoebae as well as from two multicellular developmental stages was sequenced. Computational analyses combined with experimental data reveal the expression of miRNAs, several of them exhibiting distinct expression patterns during development. To our knowledge, this is the first report of miRNAs in the Amoebozoa supergroup. We also show that overexpressed miRNA precursors generate miRNAs and, in most cases, miRNA* sequences, whose biogenesis is dependent on the Dicer-like protein DrnB, further supporting the presence of miRNAs in D. discoideum. In addition, we find miRNAs processed from hairpin structures originating from an intron as well as from a class of repetitive elements. We believe that these repetitive elements are sources for newly evolved miRNAs.

Cooperation induces other cooperation: Fruiting bodies promote the evolution of macrocysts in Dictyostelium discoideum.

Science.gov (United States)

Shibasaki, Shota; Shirokawa, Yuka; Shimada, Masakazu

2017-05-21

Biological studies of the evolution of cooperation are challenging because this process is vulnerable to cheating. Many mechanisms, including kin discrimination, spatial structure, or by-products of self-interested behaviors, can explain this evolution. Here we propose that the evolution of cooperation can be induced by other cooperation. To test this idea, we used a model organism Dictyostelium discoideum because it exhibits two cooperative dormant phases, the fruiting body and the macrocyst. In both phases, the same chemoattractant, cyclic AMP (cAMP), is used to collect cells. This common feature led us to hypothesize that the evolution of macrocyst formation would be induced by coexistence with fruiting bodies. Before forming a mathematical model, we confirmed that macrocysts coexisted with fruiting bodies, at least under laboratory conditions. Next, we analyzed our evolutionary game theory-based model to investigate whether coexistence with fruiting bodies would stabilize macrocyst formation. The model suggests that macrocyst formation represents an evolutionarily stable strategy and a global invader strategy under this coexistence, but is unstable if the model ignores the fruiting body formation. This result indicates that the evolution of macrocyst formation and maintenance is attributable to coexistence with fruiting bodies. Therefore, macrocyst evolution can be considered as an example of evolution of cooperation induced by other cooperation. Copyright © 2017 Elsevier Ltd. All rights reserved.

Partial genetic suppression of a loss-of-function mutant of the neuronal ceroid lipofuscinosis-associated protease TPP1 in Dictyostelium discoideum

Directory of Open Access Journals (Sweden)

Jonathan E. Phillips

2015-02-01

Full Text Available Neuronal ceroid lipofuscinosis (NCL is the most common childhood-onset neurodegenerative disease. NCL is inevitably fatal, and there is currently no treatment available. Children with NCL show a progressive decline in movement, vision and mental abilities, and an accumulation of autofluorescent deposits in neurons and other cell types. Late-infantile NCL is caused by mutations in the lysosomal protease tripeptidyl peptidase 1 (TPP1. TPP1 cleaves tripeptides from the N-terminus of proteins in vitro, but little is known about the physiological function of TPP1. TPP1 shows wide conservation in vertebrates but it is not found in Drosophila, Caenorhabditis elegans or Saccharomyces cerevisiae. Here, we characterize ddTpp1, a TPP1 ortholog present in the social amoeba Dictyostelium discoideum. Lysates from cells lacking ddTpp1 show a reduced but not abolished ability to cleave a TPP1 substrate, suggesting that other Dictyostelium enzymes can perform this cleavage. ddTpp1 and human TPP1 localize to the lysosome in Dictyostelium, indicating conserved function and trafficking. Cells that lack ddTpp1 show precocious multicellular development and a reduced ability to form spores during development. When cultured in autophagy-stimulating conditions, cells lacking ddTpp1 rapidly decrease in size and are less viable than wild-type cells, suggesting that one function of ddTpp1 could be to limit autophagy. Cells that lack ddTpp1 exhibit strongly impaired development in the presence of the lysosome-perturbing drug chloroquine, and this phenotype can be suppressed through a secondary mutation in the gene that we name suppressor of tpp1− A (stpA, which encodes a protein with some similarity to mammalian oxysterol-binding proteins (OSBPs. Taken together, these results suggest that targeting specific proteins could be a viable way to suppress the effects of loss of TPP1 function.

Dicty_cDB: [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VF (Link to library) VFC115 (Link to dictyBase) - - - Contig-U16382-1 VFC115E (Link...) Clone ID VFC115 (Link to dictyBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U16382-1 Ori...ant alignments: (bits) Value N X03281 |X03281.1 Dictyostelium discoideum gene for actin A8. 954 0.0 7 AC116957 |AC116957.2 Dict...X4, complete sequence. 906 0.0 9 M14146 |M14146.1 D.discoideum actin 15 gene, complete cds. 890 0.0 7 AC115579 |AC115579.2 Dict...0.0 7 U25660 |U25660.1 Dictyostelium discoideum actin gene, partial cds. 866 0.0

Dicty_cDB: [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VF (Link to library) VFB886 (Link to dictyBase) - - - Contig-U16382-1 VFB886E (Link...) Clone ID VFB886 (Link to dictyBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U16382-1 Ori...t alignments: (bits) Value N X03281 |X03281.1 Dictyostelium discoideum gene for actin A8. 2234 0.0 1 AC116957 |AC116957.2 Dict...4, complete sequence. 2107 0.0 3 M14146 |M14146.1 D.discoideum actin 15 gene, complete cds. 2034 0.0 1 AC115579 |AC115579.2 Dict...4 0.0 2 AC116986 |AC116986.2 Dictyostelium discoideum chromosome 2 map 2234041-25

Naringenin is a novel inhibitor of Dictyostelium cell proliferation and cell migration

International Nuclear Information System (INIS)

Russ, Misty; Martinez, Raquel; Ali, Hind; Steimle, Paul A.

2006-01-01

Naringenin is a flavanone compound that alters critical cellular processes such as cell multiplication, glucose uptake, and mitochondrial activity. In this study, we used the social amoeba, Dictyostelium discoideum, as a model system for examining the cellular processes and signaling pathways affected by naringenin. We found that naringenin inhibited Dictyostelium cell division in a dose-dependent manner (IC 5 ∼ 20 μM). Assays of Dictyostelium chemotaxis and multicellular development revealed that naringenin possesses a previously unrecognized ability to suppress amoeboid cell motility. We also found that naringenin, which is known to inhibit phosphatidylinositol 3-kinase activity, had no apparent effect on phosphatidylinositol 3,4,5-trisphosphate synthesis in live Dictyostelium cells; suggesting that this compound suppresses cell growth and migration via alternative signaling pathways. In another context, the discoveries described here highlight the value of using the Dictyostelium model system for identifying and characterizing the mechanisms by which naringenin, and related compounds, exert their effects on eukaryotic cells

Dicty_cDB: CFE853 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available CF (Link to library) CFE853 (Link to dictyBase) - - - Contig-U16381-1 CFE853F (Link... to Original site) CFE853F 109 - - - - - - Show CFE853 Library CF (Link to library) Clone ID CFE853 (Link to dict...yBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U16381-1 Original site URL http://dict...uences producing significant alignments: (bits) Value N X51892 |X51892.1 Dictyost...elium discoideum SP60 gene for spore coat protein. 80 6e-21 2 X52105 |X52105.1 Dictyostelium discoideum SP60

Dicty_cDB: AFK740 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available AF (Link to library) AFK740 (Link to dictyBase) - - - Contig-U16381-1 AFK740F (Link... to Original site) AFK740F 107 - - - - - - Show AFK740 Library AF (Link to library) Clone ID AFK740 (Link to dict...yBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U16381-1 Original site URL http://dict...es producing significant alignments: (bits) Value N X51892 |X51892.1 Dictyosteliu...m discoideum SP60 gene for spore coat protein. 80 6e-21 2 X52105 |X52105.1 Dictyostelium discoideum SP60 gen

Uncovering a role for the tail of the Dictyostelium discoideum SadA protein in cell-substrate adhesion.

Science.gov (United States)

Kowal, Anthony S; Chisholm, Rex L

2011-05-01

Previous work from our laboratory showed that the Dictyostelium discoideum SadA protein plays a central role in cell-substrate adhesion. SadA null cells exhibit a loss of adhesion, a disrupted actin cytoskeleton, and a cytokinesis defect. How SadA mediates these phenotypes is unknown. This work addresses the mechanism of SadA function, demonstrating an important role for the C-terminal cytoplasmic tail in SadA function. We found that a SadA tailless mutant was unable to rescue the sadA adhesion deficiency, and overexpression of the SadA tail domain reduced adhesion in wild-type cells. We also show that SadA is closely associated with the actin cytoskeleton. Mutagenesis studies suggested that four serine residues in the tail, S924/S925 and S940/S941, may regulate association of SadA with the actin cytoskeleton. Glutathione S-transferase pull-down assays identified at least one likely interaction partner of the SadA tail, cortexillin I, a known actin bundling protein. Thus, our data demonstrate an important role for the carboxy-terminal cytoplasmic tail in SadA function and strongly suggest that a phosphorylation event in this tail regulates an interaction with cortexillin I. Based on our data, we propose a model for the function of SadA.

The rate and effects of spontaneous mutation on fitness traits in the social amoeba, Dictyostelium discoideum.

Science.gov (United States)

Hall, David W; Fox, Sara; Kuzdzal-Fick, Jennie J; Strassmann, Joan E; Queller, David C

2013-07-08

We performed a mutation accumulation (MA) experiment in the social amoeba Dictyostelium discoideum to estimate the rate and distribution of effects of spontaneous mutations affecting eight putative fitness traits. We found that the per-generation mutation rate for most fitness components is 0.0019 mutations per haploid genome per generation or larger. This rate is an order of magnitude higher than estimates for fitness components in the unicellular eukaryote Saccharomyces cerevisiae, even though the base-pair substitution rate is two orders of magnitude lower. The high rate of fitness-altering mutations observed in this species may be partially explained by a large mutational target relative to S. cerevisiae. Fitness-altering mutations also may occur primarily at simple sequence repeats, which are common throughout the genome, including in coding regions, and may represent a target that is particularly likely to give fitness effects upon mutation. The majority of mutations had deleterious effects on fitness, but there was evidence for a substantial fraction, up to 40%, being beneficial for some of the putative fitness traits. Competitive ability within the multicellular slug appears to be under weak directional selection, perhaps reflecting the fact that slugs are sometimes, but not often, comprised of multiple clones in nature. Evidence for pleiotropy among fitness components across MA lines was absent, suggesting that mutations tend to act on single fitness components.

Systematic Analysis of γ-Aminobutyric Acid (GABA) Metabolism and Function in the Social Amoeba Dictyostelium discoideum*

Science.gov (United States)

Wu, Yuantai; Janetopoulos, Chris

2013-01-01

While GABA has been suggested to regulate spore encapsulation in the social amoeba Dictyostelium discoideum, the metabolic profile and other potential functions of GABA during development remain unclear. In this study, we investigated the homeostasis of GABA metabolism by disrupting genes related to GABA metabolism and signaling. Extracellular levels of GABA are tightly regulated during early development, and GABA is generated by the glutamate decarboxylase, GadB, during growth and in early development. However, overexpression of the prespore-specific homologue, GadA, in the presence of GadB reduces production of extracellular GABA. Perturbation of extracellular GABA levels delays the process of aggregation. Cytosolic GABA is degraded by the GABA transaminase, GabT, in the mitochondria. Disruption of a putative vesicular GABA transporter (vGAT) homologue DdvGAT reduces secreted GABA. We identified the GABAB receptor-like family member GrlB as the major GABA receptor during early development, and either disruption or overexpression of GrlB delays aggregation. This delay is likely the result of an abolished pre-starvation response and late expression of several “early” developmental genes. Distinct genes are employed for GABA generation during sporulation. During sporulation, GadA alone is required for generating GABA and DdvGAT is likely responsible for GABA secretion. GrlE but not GrlB is the GABA receptor during late development. PMID:23548898

Dicty_cDB: VHJ195 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available -B/AFN547Q.Seq.d/ 1342 0.0 SLE172 (SLE172Q) /CSM/SL/SLE1-C/SLE172Q.Seq.d/ 1102 0.0 SLD492 (SLD492Q) /CSM/SL/SLD4-D/SLD492...8502 ) Dictyostelium discoideum cDNA clone:dda28m11, 5' ... 839 0.0 2 ( AU052538 ) Dictyostelium discoideum slug cDNA, clone SLD492...2597 ) Dictyostelium discoideum slug cDNA, clone SLD569. 113 2e-20 1 ( AU061182 ) Dictyostelium discoideum slug cDNA, clone SLD492

Dicty_cDB: CFH809 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available CF (Link to library) CFH809 (Link to dictyBase) - - - Contig-U16381-1 - (Link to Or...iginal site) CFH809F 135 - - - - - - Show CFH809 Library CF (Link to library) Clone ID CFH809 (Link to dicty...Base) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U16381-1 Original site URL http://dictycdb.b...ce. 82 2e-29 3 X51892 |X51892.1 Dictyostelium discoideum SP60 gene for spore coat protein. 82 5e-29 2 X52105 |X52105.1 Dict...yostelium discoideum SP60 gene for spore coat protein. 80 9e-27 2 AC116977 |AC116977.2 Dict

Functional similarities between the dictyostelium protein AprA and the human protein dipeptidyl-peptidase IV.

Science.gov (United States)

Herlihy, Sarah E; Tang, Yu; Phillips, Jonathan E; Gomer, Richard H

2017-03-01

Autocrine proliferation repressor protein A (AprA) is a protein secreted by Dictyostelium discoideum cells. Although there is very little sequence similarity between AprA and any human protein, AprA has a predicted structural similarity to the human protein dipeptidyl peptidase IV (DPPIV). AprA is a chemorepellent for Dictyostelium cells, and DPPIV is a chemorepellent for neutrophils. This led us to investigate if AprA and DPPIV have additional functional similarities. We find that like AprA, DPPIV is a chemorepellent for, and inhibits the proliferation of, D. discoideum cells, and that AprA binds some DPPIV binding partners such as fibronectin. Conversely, rAprA has DPPIV-like protease activity. These results indicate a functional similarity between two eukaryotic chemorepellent proteins with very little sequence similarity, and emphasize the usefulness of using a predicted protein structure to search a protein structure database, in addition to searching for proteins with similar sequences. © 2016 The Protein Society.

Functional similarities between the dictyostelium protein AprA and the human protein dipeptidyl‐peptidase IV

Science.gov (United States)

Herlihy, Sarah E.; Tang, Yu; Phillips, Jonathan E.

2017-01-01

Abstract Autocrine proliferation repressor protein A (AprA) is a protein secreted by Dictyostelium discoideum cells. Although there is very little sequence similarity between AprA and any human protein, AprA has a predicted structural similarity to the human protein dipeptidyl peptidase IV (DPPIV). AprA is a chemorepellent for Dictyostelium cells, and DPPIV is a chemorepellent for neutrophils. This led us to investigate if AprA and DPPIV have additional functional similarities. We find that like AprA, DPPIV is a chemorepellent for, and inhibits the proliferation of, D. discoideum cells, and that AprA binds some DPPIV binding partners such as fibronectin. Conversely, rAprA has DPPIV‐like protease activity. These results indicate a functional similarity between two eukaryotic chemorepellent proteins with very little sequence similarity, and emphasize the usefulness of using a predicted protein structure to search a protein structure database, in addition to searching for proteins with similar sequences. PMID:28028841

Dicty_cDB: CFH244 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available CF (Link to library) CFH244 (Link to dictyBase) - - - Contig-U16381-1 CFH244F (Link... to Original site) CFH244F 124 - - - - - - Show CFH244 Library CF (Link to library) Clone ID CFH244 (Link to dict...yBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U16381-1 Original site URL http://dict...ts) Value N AC115685 |AC115685.1 Dictyostelium discoideum chromosome 2 map 4718821-4752388 strain AX4, compl...ete sequence. 82 2e-29 3 X51892 |X51892.1 Dictyostelium discoideum SP60 gene for spore coat protein. 82 4e-29 2 X52105 |X52105.1 Dict

Differentiation-inducing factor-1 and -2 function also as modulators for Dictyostelium chemotaxis.

Directory of Open Access Journals (Sweden)

Hidekazu Kuwayama

Full Text Available BACKGROUND: In the early stages of development of the cellular slime mold Dictyostelium discoideum, chemotaxis toward cAMP plays a pivotal role in organizing discrete cells into a multicellular structure. In this process, a series of signaling molecules, such as G-protein-coupled cell surface receptors for cAMP, phosphatidylinositol metabolites, and cyclic nucleotides, function as the signal transducers for controlling dynamics of cytoskeleton. Differentiation-inducing factor-1 and -2 (DIF-1 and DIF-2 were originally identified as the factors (chlorinated alkylphenones that induce Dictyostelium stalk cell differentiation, but it remained unknown whether the DIFs had any other physiologic functions. METHODOLOGY/PRINCIPAL FINDINGS: To further elucidate the functions of DIFs, in the present study we investigated their effects on chemotaxis under various conditions. Quite interestingly, in shallow cAMP gradients, DIF-1 suppressed chemotaxis whereas DIF-2 promoted it greatly. Analyses with various mutants revealed that DIF-1 may inhibit chemotaxis, at least in part, via GbpB (a phosphodiesterase and a decrease in the intracellular cGMP concentration ([cGMP](i. DIF-2, by contrast, may enhance chemotaxis, at least in part, via RegA (another phosphodiesterase and an increase in [cGMP](i. Using null mutants for DimA and DimB, the transcription factors that are required for DIF-dependent prestalk differentiation, we also showed that the mechanisms for the modulation of chemotaxis by DIFs differ from those for the induction of cell differentiation by DIFs, at least in part. CONCLUSIONS/SIGNIFICANCE: Our findings indicate that DIF-1 and DIF-2 function as negative and positive modulators for Dictyostelium chemotaxis, respectively. To our knowledge, this is the first report in any organism of physiologic modulators (small molecules for chemotaxis having differentiation-inducing activity.

Dicty_cDB: CHD534 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available CH (Link to library) CHD534 (Link to dictyBase) - - - Contig-U15540-1 CHD534E (Link...) Clone ID CHD534 (Link to dictyBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U15540-1 Ori...nts: (bits) Value N AC114263 |AC114263.2 Dictyostelium discoideum chromosome 2 ma...p 215673-367476 strain AX4, complete sequence. 40 1e-05 6 AC117081 |AC117081.2 Dictyostelium discoideum chro...mosome 2 map 5862124-6045772 strain AX4, complete sequence. 40 2e-05 5 AJ277590 |AJ277590.1 Dictyostelium di

Dicty_cDB: CFH744 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available CF (Link to library) CFH744 (Link to dictyBase) - - - Contig-U16381-1 - (Link to Or...iginal site) CFH744F 118 - - - - - - Show CFH744 Library CF (Link to library) Clone ID CFH744 (Link to dicty...Base) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U16381-1 Original site URL http://dictycdb.b...ng significant alignments: (bits) Value N X51892 |X51892.1 Dictyostelium discoide...um SP60 gene for spore coat protein. 80 1e-23 3 AC115685 |AC115685.1 Dictyostelium discoideum chromosome 2 m

Extracellular calmodulin regulates growth and cAMP-mediated chemotaxis in Dictyostelium discoideum

International Nuclear Information System (INIS)

O’Day, Danton H.; Huber, Robert J.; Suarez, Andres

2012-01-01

Highlights: ► Extracellular calmodulin is present throughout growth and development in Dictyostelium. ► Extracellular calmodulin localizes within the ECM during development. ► Extracellular calmodulin inhibits cell proliferation and increases chemotaxis. ► Extracellular calmodulin exists in eukaryotic microbes. ► Extracellular calmodulin may be functionally as important as intracellular calmodulin. -- Abstract: The existence of extracellular calmodulin (CaM) has had a long and controversial history. CaM is a ubiquitous calcium-binding protein that has been found in every eukaryotic cell system. Calcium-free apo-CaM and Ca 2+ /CaM exert their effects by binding to and regulating the activity of CaM-binding proteins (CaMBPs). Most of the research done to date on CaM and its CaMBPs has focused on their intracellular functions. The presence of extracellular CaM is well established in a number of plants where it functions in proliferation, cell wall regeneration, gene regulation and germination. While CaM has been detected extracellularly in several animal species, including frog, rat, rabbit and human, its extracellular localization and functions are less well established. In contrast the study of extracellular CaM in eukaryotic microbes remains to be done. Here we show that CaM is constitutively expressed and secreted throughout asexual development in Dictyostelium where the presence of extracellular CaM dose-dependently inhibits cell proliferation but increases cAMP mediated chemotaxis. During development, extracellular CaM localizes within the slime sheath where it coexists with at least one CaMBP, the matricellular CaM-binding protein CyrA. Coupled with previous research, this work provides direct evidence for the existence of extracellular CaM in the Dictyostelium and provides insight into its functions in this model amoebozoan.

The Parkinson's disease-associated protein DJ-1 plays a positive nonmitochondrial role in endocytosis in Dictyostelium cells

Directory of Open Access Journals (Sweden)

Suwei Chen

2017-10-01

Full Text Available The loss of function of DJ-1 caused by mutations in DJ1 causes a form of familial Parkinson's disease (PD. However, the role of DJ-1 in healthy and in PD cells is poorly understood. Even its subcellular localization in mammalian cells is uncertain, with both cytosolic and mitochondrial locations having been reported. We show here that DJ-1 is normally located in the cytoplasm in healthy Dictyostelium discoideum cells. With its unique life cycle, straightforward genotype-phenotype relationships, experimental accessibility and genetic tractability, D. discoideum offers an attractive model to investigate the roles of PD-associated genes. Furthermore, the study of mitochondrial biology, mitochondrial genome transcription and AMP-activated protein kinase-mediated cytopathologies in mitochondrial dysfunction have been well developed in this organism. Unlike mammalian systems, Dictyostelium mitochondrial dysfunction causes a reproducible and readily assayed array of aberrant phenotypes: defective phototaxis, impaired growth, normal rates of endocytosis and characteristic defects in multicellular morphogenesis. This makes it possible to study whether the underlying cytopathological mechanisms of familial PD involve mitochondrial dysfunction. DJ-1 has a single homologue in the Dictyostelium genome. By regulating the expression level of DJ-1 in D. discoideum, we show here that in unstressed cells, DJ-1 is required for normal rates of endocytic nutrient uptake (phagocytosis and, to a lesser extent, pinocytosis and thus growth. Reduced expression of DJ-1 had no effect on phototaxis in the multicellular migratory ‘slug’ stage of the life cycle, but resulted in thickened stalks in the final fruiting bodies. This pattern of phenotypes is distinct from that observed in Dictyostelium to result from mitochondrial dyfunction. Direct measurement of mitochondrial respiratory function in intact cells revealed that DJ-1 knockdown stimulates whereas DJ-1

Regulation of Spatiotemporal Patterns by Biological Variability: General Principles and Applications to Dictyostelium discoideum.

Directory of Open Access Journals (Sweden)

Miriam Grace

2015-11-01

Full Text Available Spatiotemporal patterns often emerge from local interactions in a self-organizing fashion. In biology, the resulting patterns are also subject to the influence of the systematic differences between the system's constituents (biological variability. This regulation of spatiotemporal patterns by biological variability is the topic of our review. We discuss several examples of correlations between cell properties and the self-organized spatiotemporal patterns, together with their relevance for biology. Our guiding, illustrative example will be spiral waves of cAMP in a colony of Dictyostelium discoideum cells. Analogous processes take place in diverse situations (such as cardiac tissue, where spiral waves occur in potentially fatal ventricular fibrillation so a deeper understanding of this additional layer of self-organized pattern formation would be beneficial to a wide range of applications. One of the most striking differences between pattern-forming systems in physics or chemistry and those in biology is the potential importance of variability. In the former, system components are essentially identical with random fluctuations determining the details of the self-organization process and the resulting patterns. In biology, due to variability, the properties of potentially very few cells can have a driving influence on the resulting asymptotic collective state of the colony. Variability is one means of implementing a few-element control on the collective mode. Regulatory architectures, parameters of signaling cascades, and properties of structure formation processes can be "reverse-engineered" from observed spatiotemporal patterns, as different types of regulation and forms of interactions between the constituents can lead to markedly different correlations. The power of this biology-inspired view of pattern formation lies in building a bridge between two scales: the patterns as a collective state of a very large number of cells on the one hand

Determination of inositol 1,4,5-trisphosphate levels in Dictyostelium by isotope dilution assay

International Nuclear Information System (INIS)

Van Haastert, P.J.

1989-01-01

A commercial isotope dilution assay was used for the determination of Ins(1,4,5)P3 levels in the microorganism Dictyostelium discoideum. Cross-reactivity in the assay was detected with extracts from cells and the medium. The compound which induced this cross-reactivity was tentatively identified as Ins(1,4,5)P3 by (i) codegradation with authentic [ 32 P]Ins(1,4,5)P3 by three specific Ins(1,4,5)P3 phosphatases, and (ii) co-chromatography with authentic [ 32 P]Ins(1,4,5)P3 on HPLC columns. The cellular concentration was estimated as 165 +/- 42 pmol/10(8) cells, yielding a mean intracellular Ins(1,4,5)P3 concentration of 3.3 microM. Dictyostelium cells secrete large amounts of Ins(1,4,5)P3 at a rate of about 10% of the cellular content per minute, yielding about 0.13 microM extracellular Ins(1,4,5)P3 after 15 min in a suspension of 10(8) cells/ml. The chemoattractant cAMP induced a transient increase of the Ins(1,4,5)P3 concentration; the data suggest an intracacellular rise from 3.3 to 5.5 microM with a maximum at 6 s after stimulation

Dicty_cDB: Contig-U13443-1 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available lignments: (bits) Value N ( AF305060 ) Dictyostelium discoideum Wiscott-Aldrich syndrome... 529 0.0 10 ( BJ3... AF305060 ) Dictyostelium discoideum Wiscott-Aldrich syndrome protein (wasA) gene...icant alignments: (bits) Value AF305060_1( AF305060 |pid:none) Dictyostelium discoideum Wiscott...0_1( AF305060 |pid:none) Dictyostelium discoideum Wiscott-Aldrich syndrome protein (wasA) gene, complete cds

The ROCO kinase QkgA is necessary for proliferation inhibition by autocrine signals in Dictyostelium discoideum.

Science.gov (United States)

Phillips, Jonathan E; Gomer, Richard H

2010-10-01

AprA and CfaD are secreted proteins that function as autocrine signals to inhibit cell proliferation in Dictyostelium discoideum. Cells lacking AprA or CfaD proliferate rapidly, and adding AprA or CfaD to cells slows proliferation. Cells lacking the ROCO kinase QkgA proliferate rapidly, with a doubling time 83% of that of the wild type, and overexpression of a QkgA-green fluorescent protein (GFP) fusion protein slows cell proliferation. We found that qkgA(-) cells accumulate normal levels of extracellular AprA and CfaD. Exogenous AprA or CfaD does not slow the proliferation of cells lacking qkgA, and expression of QkgA-GFP in qkgA(-) cells rescues this insensitivity. Like cells lacking AprA or CfaD, cells lacking QkgA tend to be multinucleate, accumulate nuclei rapidly, and show a mass and protein accumulation per nucleus like those of the wild type, suggesting that QkgA negatively regulates proliferation but not growth. Despite their rapid proliferation, cells lacking AprA, CfaD, or QkgA expand as a colony on bacteria less rapidly than the wild type. Unlike AprA and CfaD, QkgA does not affect spore viability following multicellular development. Together, these results indicate that QkgA is necessary for proliferation inhibition by AprA and CfaD, that QkgA mediates some but not all of the effects of AprA and CfaD, and that QkgA may function downstream of these proteins in a signal transduction pathway regulating proliferation.

Dicty_cDB: AFM869 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available Dictyostelium discoideum slug cDNA, clone SLH341. 404 e-128 3 ( AF305060 ) Dictyostelium discoideum Wiscott-...ucing significant alignments: (bits) Value AF305060_1( AF305060 |pid:none) Dictyostelium discoideum Wiscott-

Dicty_cDB: [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VF (Link to library) VFB593 (Link to dictyBase) - - - Contig-U02438-1 VFB593E (Link...) Clone ID VFB593 (Link to dictyBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U02438-1 Ori...0.009 6 AC116986 |AC116986.2 Dictyostelium discoideum chromosome 2 map 2234041-25...sequence. 46 0.031 2 AC115577 |AC115577.2 Dictyostelium discoideum chromosome 2 m...ap 4657875-4914984 strain AX4, complete sequence. 34 0.051 14 AC116960 |AC116960.2 Dictyostelium discoideum

Uncovering a Role for the Tail of the Dictyostelium discoideum SadA Protein in Cell-Substrate Adhesion ▿ †

Science.gov (United States)

Kowal, Anthony S.; Chisholm, Rex L.

2011-01-01

Previous work from our laboratory showed that the Dictyostelium discoideum SadA protein plays a central role in cell-substrate adhesion. SadA null cells exhibit a loss of adhesion, a disrupted actin cytoskeleton, and a cytokinesis defect. How SadA mediates these phenotypes is unknown. This work addresses the mechanism of SadA function, demonstrating an important role for the C-terminal cytoplasmic tail in SadA function. We found that a SadA tailless mutant was unable to rescue the sadA adhesion deficiency, and overexpression of the SadA tail domain reduced adhesion in wild-type cells. We also show that SadA is closely associated with the actin cytoskeleton. Mutagenesis studies suggested that four serine residues in the tail, S924/S925 and S940/S941, may regulate association of SadA with the actin cytoskeleton. Glutathione S-transferase pull-down assays identified at least one likely interaction partner of the SadA tail, cortexillin I, a known actin bundling protein. Thus, our data demonstrate an important role for the carboxy-terminal cytoplasmic tail in SadA function and strongly suggest that a phosphorylation event in this tail regulates an interaction with cortexillin I. Based on our data, we propose a model for the function of SadA. PMID:21441344

NCBI nr-aa BLAST: CBRC-DDIS-01-0111 [SEVENS

Lifescience Database Archive (English)

Full Text Available CBRC-DDIS-01-0111 ref|XP_636760.1| alkaline dihydroceramidase [Dictyostelium discoi...deum AX4] gb|AAQ98884.1| alkaline dihydroceramidase [Dictyostelium discoideum] gb|EAL63276.1| alkaline dihydroceramidase [Dictyostelium discoideum AX4] XP_636760.1 7e-59 45% ...

Structure, dynamics and folding of an immunoglobulin domain of the gelation factor (ABP-120) from Dictyostelium discoideum.

Science.gov (United States)

Hsu, Shang-Te Danny; Cabrita, Lisa D; Fucini, Paola; Dobson, Christopher M; Christodoulou, John

2009-05-15

We have carried out a detailed structural and dynamical characterisation of the isolated fifth repeat of the gelation factor (ABP-120) from Dictyostelium discoideum (ddFLN5) by NMR spectroscopy to provide a basis for studies of co-translational folding on the ribosome of this immunoglobulin-like domain. The isolated ddFLN5 can fold autonomously in solution into a structure that resembles very closely the crystal structure of the domain in a construct in which the adjacent sixth repeat (ddFLN6) is covalently linked to its C-terminus in tandem but deviates locally from a second crystal structure in which ddFLN5 is flanked by ddFLN4 and ddFLN6 at both N- and C-termini. Conformational fluctuations were observed via (15)N relaxation methods and are primarily localised in the interstrand loops that encompass the C-terminal hemisphere. These fluctuations are distinct in location from the region where line broadening is observed in ddFLN5 when attached to the ribosome as part of a nascent chain. This observation supports the conclusion that the broadening is associated with interactions with the ribosome surface [Hsu, S. T. D., Fucini, P., Cabrita, L. D., Launay, H., Dobson, C. M. & Christodoulou, J. (2007). Structure and dynamics of a ribosome-bound nascent chain by NMR spectroscopy. Proc. Natl. Acad. Sci. USA, 104, 16516-16521]. The unfolding of ddFLN5 induced by high concentrations of urea shows a low population of a folding intermediate, as inferred from an intensity-based analysis, a finding that differs from that of ddFLN5 as a ribosome-bound nascent chain. These results suggest that interesting differences in detail may exist between the structure of the domain in isolation and when linked to the ribosome and between protein folding in vitro and the folding of a nascent chain as it emerges from the ribosome.

NCBI nr-aa BLAST: CBRC-DDIS-01-0132 [SEVENS

Lifescience Database Archive (English)

Full Text Available CBRC-DDIS-01-0132 ref|XP_646256.1| cellulose synthase [Dictyostelium discoideum AX4...] gb|AAF00200.1|AF163835_1 cellulose synthase [Dictyostelium discoideum] gb|EAL71912.1| cellulose synthase [Dictyostelium discoideum AX4] XP_646256.1 0.0 99% ...

Dicty_cDB: [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VF (Link to library) VFB489 (Link to dictyBase) - - - Contig-U16382-1 VFB489P (Link... to Original site) VFB489F 178 VFB489Z 501 VFB489P 679 - - Show VFB489 Library VF (Link to library) Clone ID VFB489 (Link to dict...yBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U16382-1 Original site URL http://dict...: (bits) Value N AC116986 |AC116986.2 Dictyostelium discoideum chromosome 2 map 2...e cds. 783 0.0 3 AC115579 |AC115579.2 Dictyostelium discoideum chromosome 2 map 4915084-5005461 strain AX4,

Dicty_cDB: [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VF (Link to library) VFC187 (Link to dictyBase) - - - Contig-U12289-1 VFC187P (Link... to Original site) VFC187F 411 VFC187Z 678 VFC187P 1089 - - Show VFC187 Library VF (Link to library) Clone ID VFC187 (Link to dict...yBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U12289-1 Original site URL http://dict... N AC117176 |AC117176.2 Dictyostelium discoideum chromosome 2 map 5018074-5200947... strain AX4, complete sequence. 36 0.008 12 AC114263 |AC114263.2 Dictyostelium discoideum chromosome 2 map 2

Dicty_cDB: [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VF (Link to library) VFB837 (Link to dictyBase) - - - Contig-U14985-1 VFB837P (Link... to Original site) VFB837F 606 VFB837Z 689 VFB837P 1295 - - Show VFB837 Library VF (Link to library) Clone ID VFB837 (Link to dict...yBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U14985-1 Original site URL http://dict... 13 AC116986 |AC116986.2 Dictyostelium discoideum chromosome 2 map 2234041-256737...0 strain AX4, complete sequence. 38 2e-05 15 AC116984 |AC116984.2 Dictyostelium discoideum chromosome 2 map

Dicty_cDB: SFJ736 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available 060.1 Dictyostelium discoideum Wiscott-Aldrich syndrome protein (wasA) gene, complete cds. 470 e-129 2 BQ923...1( AF305060 |pid:none) Dictyostelium discoideum Wiscott-A... 81 1e-14 AC117076_18

Properties of a non-bioactive fluorescent derivative of differentiation-inducing factor-3, an anti-tumor agent found in Dictyostelium discoideum

Directory of Open Access Journals (Sweden)

Yuzuru Kubohara

2014-03-01

Full Text Available Differentiation-inducing factor-3 (DIF-3, found in the cellular slime mold Dictyostelium discoideum, and its derivatives, such as butoxy-DIF-3 (Bu-DIF-3, are potent anti-tumor agents. To investigate the activity of DIF-like molecules in tumor cells, we recently synthesized a green fluorescent DIF-3 derivative, BODIPY-DIF-3G, and analyzed its bioactivity and cellular localization. In this study, we synthesized a red (orange fluorescent DIF-3 derivative, BODIPY-DIF-3R, and compared the cellular localization and bioactivities of the two BODIPY-DIF-3s in HeLa human cervical cancer cells. Both fluorescent compounds penetrated the extracellular membrane within 0.5 h and localized mainly to the mitochondria. In formalin-fixed cells, the two BODIPY-DIF-3s also localized to the mitochondria, indicating that the BODIPY-DIF-3s were incorporated into mitochondria independently of the mitochondrial membrane potential. After treatment for 3 days, BODIPY-DIF-3G, but not BODIPY-DIF-3R, induced mitochondrial swelling and suppressed cell proliferation. Interestingly, the swollen mitochondria were stainable with BODIPY-DIF-3G but not with BODIPY-DIF-3R. When added to isolated mitochondria in vitro, BODIPY-DIF-3G increased dose-dependently the rate of O2 consumption, but BODIPY-DIF-3R did not. These results suggest that the bioactive BODIPY-DIF-3G suppresses cell proliferation, at least in part, by altering mitochondrial activity, whereas the non-bioactive BODIPY-DIF-3R localizes to the mitochondria but does not affect mitochondrial activity or cell proliferation.

Dicty_cDB: VSH207 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VS (Link to library) VSH207 (Link to dictyBase) - - - Contig-U12548-1 VSH207P (Link... to Original site) VSH207F 228 VSH207Z 107 VSH207P 335 - - Show VSH207 Library VS (Link to library) Clone ID VSH207 (Link to dict...yBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U12548-1 Original site URL http://dict...ents: (bits) Value N ( AU267072 ) Dictyostelium discoideum vegetative cDNA clone:VS... 206 2e-58 2 ( AC116982 ) Dict...yostelium discoideum chromosome 2 map 3622643... 206 4e-49 1 ( AU267073 ) Dict

Dicty_cDB: [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VF (Link to library) VFB788 (Link to dictyBase) - - - Contig-U14924-1 VFB788P (Link... to Original site) VFB788F 158 VFB788Z 768 VFB788P 926 - - Show VFB788 Library VF (Link to library) Clone ID VFB788 (Link to dict...yBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U14924-1 Original site URL http://dict... (bits) Value N AC115592 |AC115592.2 Dictyostelium discoideum chromosome 2 map 1-...2_6( AC115592 |pid:none) Dictyostelium discoideum chromosom... 520 e-146 CU459003_2449( CU459003 |pid:none)

Dicty_cDB: Contig-U12991-1 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available ( AF272150 ) Dictyostelium discoideum deliriumA (dlrA) gene, c... 2022 0.0 3 ( BJ...39594 ) TT1EP48TV Tetrahymena thermophila SB210 cDNA libr... 38 10.0 2 >( AF272150 ) Dictyostelium discoideum delirium

Mitochondria are the target organelle of differentiation-inducing factor-3, an anti-tumor agent isolated from Dictyostelium discoideum [corrected].

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Yuzuru Kubohara

Full Text Available Differentiation-inducing factor-3 (DIF-3, found in the cellular slime mold Dictyostelium discoideum, and its derivatives such as butoxy-DIF-3 (Bu-DIF-3 are potent anti-tumor agents. However, the precise mechanisms underlying the actions of DIF-3 remain to be elucidated. In this study, we synthesized a green fluorescent derivative of DIF-3, BODIPY-DIF-3, and a control fluorescent compound, Bu-BODIPY (butyl-BODIPY, and investigated how DIF-like molecules behave in human cervical cancer HeLa cells by using both fluorescence and electron microscopy. BODIPY-DIF-3 at 5-20 µ M suppressed cell growth in a dose-dependent manner, whereas Bu-BODIPY had minimal effect on cell growth. When cells were incubated with BODIPY-DIF-3 at 20 µM, it penetrated cell membranes within 0.5 h and localized mainly in mitochondria, while Bu-BODIPY did not stain the cells. Exposure of cells for 1-3 days to DIF-3, Bu-DIF-3, BODIPY-DIF-3, or CCCP (a mitochondrial uncoupler induced substantial mitochondrial swelling, suppressing cell growth. When added to isolated mitochondria, DIF-3, Bu-DIF-3, and BOIDPY-DIF-3, like CCCP, dose-dependently promoted the rate of oxygen consumption, but Bu-BODIPY did not. Our results suggest that these bioactive DIF-like molecules suppress cell growth, at least in part, by disturbing mitochondrial activity. This is the first report showing the cellular localization and behavior of DIF-like molecules in mammalian tumor cells.

Dicty_cDB: SFK712 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available deum cDNA clone:dda5o08, 3' e... 404 e-125 2 ( AF305060 ) Dictyostelium discoideum Wiscott-Aldrich syndrome....ore E Sequences producing significant alignments: (bits) Value AF305060_1( AF305060 |pid:none) Dictyostelium discoideum Wiscott

Dicty_cDB: VSJ735 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available |AF305060.1 Dictyostelium discoideum Wiscott-Aldrich syndrome protein (wasA) gene, complete cds. 436 0.0 5 A... AF305060 |pid:none) Dictyostelium discoideum Wiscott-A... 214 2e-54 BC087802_1( BC087802 |pid:none) Xenopus

A comparative sequence analysis reveals a common GBD/FH3-FH1-FH2-DAD architecture in formins from Dictyostelium, fungi and metazoa

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Uyeda Taro QP

2005-03-01

Full Text Available Abstract Background Formins are multidomain proteins defined by a conserved FH2 (formin homology 2 domain with actin nucleation activity preceded by a proline-rich FH1 (formin homology 1 domain. Formins act as profilin-modulated processive actin nucleators conserved throughout a wide range of eukaryotes. Results We present a detailed sequence analysis of the 10 formins (ForA to J identified in the genome of the social amoeba Dictyostelium discoideum. With the exception of ForI and ForC all other formins conform to the domain structure GBD/FH3-FH1-FH2-DAD, where DAD is the Diaphanous autoinhibition domain and GBD/FH3 is the Rho GTPase-binding domain/formin homology 3 domain that we propose to represent a single domain. ForC lacks a FH1 domain, ForI lacks recognizable GBD/FH3 and DAD domains and ForA, E and J have additional unique domains. To establish the relationship between formins of Dictyostelium and other organisms we constructed a phylogenetic tree based on the alignment of FH2 domains. Real-time PCR was used to study the expression pattern of formin genes. Expression of forC, D, I and J increased during transition to multi-cellular stages, while the rest of genes displayed less marked developmental variations. During sexual development, expression of forH and forI displayed a significant increase in fusion competent cells. Conclusion Our analysis allows some preliminary insight into the functionality of Dictyostelium formins: all isoforms might display actin nucleation activity and, with the exception of ForI, might also be susceptible to autoinhibition and to regulation by Rho GTPases. The architecture GBD/FH3-FH1-FH2-DAD appears common to almost all Dictyostelium, fungal and metazoan formins, for which we propose the denomination of conventional formins, and implies a common regulatory mechanism.

A comparative sequence analysis reveals a common GBD/FH3-FH1-FH2-DAD architecture in formins from Dictyostelium, fungi and metazoa.

Science.gov (United States)

Rivero, Francisco; Muramoto, Tetsuya; Meyer, Ann-Kathrin; Urushihara, Hideko; Uyeda, Taro Q P; Kitayama, Chikako

2005-03-01

Formins are multidomain proteins defined by a conserved FH2 (formin homology 2) domain with actin nucleation activity preceded by a proline-rich FH1 (formin homology 1) domain. Formins act as profilin-modulated processive actin nucleators conserved throughout a wide range of eukaryotes. We present a detailed sequence analysis of the 10 formins (ForA to J) identified in the genome of the social amoeba Dictyostelium discoideum. With the exception of ForI and ForC all other formins conform to the domain structure GBD/FH3-FH1-FH2-DAD, where DAD is the Diaphanous autoinhibition domain and GBD/FH3 is the Rho GTPase-binding domain/formin homology 3 domain that we propose to represent a single domain. ForC lacks a FH1 domain, ForI lacks recognizable GBD/FH3 and DAD domains and ForA, E and J have additional unique domains. To establish the relationship between formins of Dictyostelium and other organisms we constructed a phylogenetic tree based on the alignment of FH2 domains. Real-time PCR was used to study the expression pattern of formin genes. Expression of forC, D, I and J increased during transition to multi-cellular stages, while the rest of genes displayed less marked developmental variations. During sexual development, expression of forH and forI displayed a significant increase in fusion competent cells. Our analysis allows some preliminary insight into the functionality of Dictyostelium formins: all isoforms might display actin nucleation activity and, with the exception of ForI, might also be susceptible to autoinhibition and to regulation by Rho GTPases. The architecture GBD/FH3-FH1-FH2-DAD appears common to almost all Dictyostelium, fungal and metazoan formins, for which we propose the denomination of conventional formins, and implies a common regulatory mechanism.

Heteromeric p97/p97R155C complexes induce dominant negative changes in wild-type and autophagy 9-deficient Dictyostelium strains.

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Khalid Arhzaouy

Full Text Available Heterozygous mutations in the human VCP (p97 gene cause autosomal-dominant IBMPFD (inclusion body myopathy with early onset Paget's disease of bone and frontotemporal dementia, ALS14 (amyotrophic lateral sclerosis with or without frontotemporal dementia and HSP (hereditary spastic paraplegia. Most prevalent is the R155C point mutation. We studied the function of p97 in the social amoeba Dictyostelium discoideum and have generated strains that ectopically express wild-type (p97 or mutant p97 (p97(R155C fused to RFP in AX2 wild-type and autophagy 9 knock-out (ATG9(KO cells. Native gel electrophoresis showed that both p97 and p97(R155C assemble into hexamers. Co-immunoprecipitation studies revealed that endogenous p97 and p97(R155C-RFP form heteromers. The mutant strains displayed changes in cell growth, phototaxis, development, proteasomal activity, ubiquitinylated proteins, and ATG8(LC3 indicating mis-regulation of multiple essential cellular processes. Additionally, immunofluorescence analysis revealed an increase of protein aggregates in ATG9(KO/p97(R155C-RFP and ATG9(KO cells. They were positive for ubiquitin in both strains, however, solely immunoreactive for p97 in the ATG9(KO mutant. A major finding is that the expression of p97(R155C-RFP in the ATG9(KO strain partially or fully rescued the pleiotropic phenotype. We also observed dose-dependent effects of p97 on several cellular processes. Based on findings in the single versus the double mutants we propose a novel mode of p97 interaction with the core autophagy protein ATG9 which is based on mutual inhibition.

G-protein-mediated interconversions of cell-surface cAMP receptors and their involvement in excitation and desensitization of guanylate cyclase in Dictyostelium discoideum

International Nuclear Information System (INIS)

van Haastert, P.J.; de Wit, R.J.; Janssens, P.M.; Kesbeke, F.; DeGoede, J.

1986-01-01

In Dictyostelium discoideum cells, extracellular cAMP induces the rapid (within 2 s) activation of guanylate cyclase, which is followed by complete desensitization after about 10 s. cAMP binding to these cells is heterogeneous, showing a subclass of fast dissociating sites coupled to adenylate cyclase (A-sites) and a subclass of slowly dissociating sites coupled to guanylate cyclase (B-sites). The kinetics of the B-sites were further investigated on a seconds time scale. Statistical analysis of the association of [ 3 H]cAMP to the B-sites and dissociation of the complex revealed that the receptor can exist in three states which interconvert according to the following scheme. cAMP binds to the BF-state (off-rate 2.5 s) which rapidly (t1/2 = 3 s) converts to the BS-state (off-rate 15 s) and subsequently (without a detectable delay) into the BSS-state (off-rate 150 s). In membranes, both the BS- and BSS-states are converted to the BF-state by GTP and GDP, suggesting the involvement of a G-protein. Densensitized cells show a 80% reduction of the formation of the BSS-state, but no reduction of the BF- or BS-state. These data are combined into a model in which the transitions of the B-sites are mediated by a G-protein; activation of the G-protein and guanylate cyclase is associated with the transition of the BS- to the BSS-state of the receptor, whereas desensitization is associated with the inhibition of this transition

A flavin-dependent halogenase catalyzes the chlorination step in the biosynthesis of Dictyostelium differentiation-inducing factor 1

OpenAIRE

Neumann, Christopher S.; Walsh, Christopher T.; Kay, Robert R.

2010-01-01

Differentiation-inducing factor 1 (DIF-1) is a polyketide-derived morphogen which drives stalk cell formation in the developmental cycle of Dictyostelium discoideum. Previous experiments demonstrated that the biosynthetic pathway proceeds via dichlorination of the precursor molecule THPH, but the enzyme responsible for this transformation has eluded characterization. Our recent studies on prokaryotic flavin-dependent halogenases and insights from the sequenced Dd genome led us to a candidate ...

The Dictyostelium Bcr/Abr-related protein DRG regulates both Rac- and Rab-dependent pathways

OpenAIRE

Knetsch, Menno L.W.; Schäfers, Nicole; Horstmann, Heinz; Manstein, Dietmar J.

2001-01-01

Dictyostelium discoideum DdRacGap1 (DRG) contains both Rho-GEF and Rho-GAP domains, a feature it shares with mammalian Bcr and Abr. To elucidate the physiological role of this multifunctional protein, we characterized the enzymatic activity of recombinant DRG fragments in vitro, created DRG-null cells, and studied the function of the protein in vivo by analysing the phenotypic changes displayed by DRG-depleted cells and DRG-null cells complemented with DRG or DRG fragments. Our results show t...

Lack of Ecological and Life History Context Can Create the Illusion of Social Interactions in Dictyostelium discoideum.

Science.gov (United States)

Martínez-García, Ricardo; Tarnita, Corina E

2016-12-01

Studies of social microbes often focus on one fitness component (reproductive success within the social complex), with little information about or attention to other stages of the life cycle or the ecological context. This can lead to paradoxical results. The life cycle of the social amoeba Dictyostelium discoideum includes a multicellular stage in which not necessarily clonal amoebae aggregate upon starvation to form a possibly chimeric (genetically heterogeneous) fruiting body made of dead stalk cells and spores. The lab-measured reproductive skew in the spores of chimeras indicates strong social antagonism that should result in low genotypic diversity, which is inconsistent with observations from nature. Two studies have suggested that this inconsistency stems from the one-dimensional assessment of fitness (spore production) and that the solution lies in tradeoffs between multiple life-history traits, e.g.: spore size versus viability; and spore-formation (via aggregation) versus staying vegetative (as non-aggregated cells). We develop an ecologically-grounded, socially-neutral model (i.e. no social interactions between genotypes) for the life cycle of social amoebae in which we theoretically explore multiple non-social life-history traits, tradeoffs and tradeoff-implementing mechanisms. We find that spore production comes at the expense of time to complete aggregation, and, depending on the experimental setup, spore size and viability. Furthermore, experimental results regarding apparent social interactions within chimeric mixes can be qualitatively recapitulated under this neutral hypothesis, without needing to invoke social interactions. This allows for simple potential resolutions to the previously paradoxical results. We conclude that the complexities of life histories, including social behavior and multicellularity, can only be understood in the appropriate multidimensional ecological context, when considering all stages of the life cycle.

Dicty_cDB: SFF823 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available gy vs DNA Score E Sequences producing significant alignments: (bits) Value N AF305060 |AF305060.1 Dictyostelium discoideum Wiscott...id:none) Dictyostelium discoideum Wiscott-A... 83 3e-15 AC117076_18( AC117076 |pid:none) Dictyostelium disco

Dicty_cDB: VFD730 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available omology vs DNA Score E Sequences producing significant alignments: (bits) Value N AF305060 |AF305060.1 Dictyostelium discoideum Wisco...otein Score E Sequences producing significant alignments: (bits) Value AF305060_1( AF305060 |pid:none) Dictyostelium discoideum Wisco...tt-Aldrich syndrome protein (wasA) gene, complete cds. 4...tt-A... 83 3e-15 AC117076_18( AC117076 |pid:none) Dictyostelium discoideum chromoso

Targets downstream of Cdk8 in Dictyostelium development

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Skelton Jason

2011-01-01

Full Text Available Abstract Background Cdk8 is a component of the mediator complex which facilitates transcription by RNA polymerase II and has been shown to play an important role in development of Dictyostelium discoideum. This eukaryote feeds as single cells but starvation triggers the formation of a multicellular organism in response to extracellular pulses of cAMP and the eventual generation of spores. Strains in which the gene encoding Cdk8 have been disrupted fail to form multicellular aggregates unless supplied with exogenous pulses of cAMP and later in development, cdk8- cells show a defect in spore production. Results Microarray analysis revealed that the cdk8- strain previously described (cdk8-HL contained genome duplications. Regeneration of the strain in a background lacking detectable gene duplication generated strains (cdk8-2 with identical defects in growth and early development, but a milder defect in spore generation, suggesting that the severity of this defect depends on the genetic background. The failure of cdk8- cells to aggregate unless rescued by exogenous pulses of cAMP is consistent with a failure to express the catalytic subunit of protein kinase A. However, overexpression of the gene encoding this protein was not sufficient to rescue the defect, suggesting that this is not the only important target for Cdk8 at this stage of development. Proteomic analysis revealed two potential targets for Cdk8 regulation, one regulated post-transcriptionally (4-hydroxyphenylpyruvate dioxygenase (HPD and one transcriptionally (short chain dehydrogenase/reductase (SDR1. Conclusions This analysis has confirmed the importance of Cdk8 at multiple stages of Dictyostelium development, although the severity of the defect in spore production depends on the genetic background. Potential targets of Cdk8-mediated gene regulation have been identified in Dictyostelium which will allow the mechanism of Cdk8 action and its role in development to be determined.

Derivatives of Dictyostelium differentiation-inducing factors inhibit lysophosphatidic acid–stimulated migration of murine osteosarcoma LM8 cells

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Kubohara, Yuzuru, E-mail: ykuboha@juntendo.ac.jp [Department of Molecular and Cellular Biology, Institute for Molecular and Cellular Regulation (IMCR), Gunma University, Maebashi 371-8512 (Japan); Department of Health Science, Juntendo University Graduate School of Health and Sports Science, Inzai 270-1695 (Japan); Komachi, Mayumi [Department of Molecular and Cellular Biology, Institute for Molecular and Cellular Regulation (IMCR), Gunma University, Maebashi 371-8512 (Japan); Department of Radiation Oncology, Gunma University Graduate School of Medicine, Maebashi 371-8511 (Japan); Homma, Yoshimi [Department of Biomolecular Science, Institute of Biomedical Sciences, Fukushima Medical University School of Medicine, Fukushima 960-1295 (Japan); Kikuchi, Haruhisa; Oshima, Yoshiteru [Laboratory of Natural Product Chemistry, Tohoku University Graduate School of Pharmaceutical Sciences, Aoba-yama, Aoba-ku, Sendai 980-8578 (Japan)

2015-08-07

Osteosarcoma is a common metastatic bone cancer that predominantly develops in children and adolescents. Metastatic osteosarcoma remains associated with a poor prognosis; therefore, more effective anti-metastatic drugs are needed. Differentiation-inducing factor-1 (DIF-1), −2, and −3 are novel lead anti-tumor agents that were originally isolated from the cellular slime mold Dictyostelium discoideum. Here we investigated the effects of a panel of DIF derivatives on lysophosphatidic acid (LPA)-induced migration of mouse osteosarcoma LM8 cells by using a Boyden chamber assay. Some DIF derivatives such as Br-DIF-1, DIF-3(+2), and Bu-DIF-3 (5–20 μM) dose-dependently suppressed LPA-induced cell migration with associated IC{sub 50} values of 5.5, 4.6, and 4.2 μM, respectively. On the other hand, the IC{sub 50} values of Br-DIF-1, DIF-3(+2), and Bu-DIF-3 versus cell proliferation were 18.5, 7.2, and 2.0 μM, respectively, in LM8 cells, and >20, 14.8, and 4.3 μM, respectively, in mouse 3T3-L1 fibroblasts (non-transformed). Together, our results demonstrate that Br-DIF-1 in particular may be a valuable tool for the analysis of cancer cell migration, and that DIF derivatives such as DIF-3(+2) and Bu-DIF-3 are promising lead anti-tumor agents for the development of therapies that suppress osteosarcoma cell proliferation, migration, and metastasis. - Highlights: • LPA induces cell migration (invasion) in murine osteosarcoma LM8 cells. • DIFs are novel lead anti-tumor agents found in Dictyostelium discoideum. • We examined the effects of DIF derivatives on LPA-induced LM8 cell migration in vitro. • Some of the DIF derivatives inhibited LPA-induced LM8 cell migration.

Dicty_cDB: [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VF (Link to library) VFC138 (Link to dictyBase) - - - Contig-U15456-1 VFC138P (Link... to Original site) VFC138F 411 VFC138Z 440 VFC138P 851 - - Show VFC138 Library VF (Link to library) Clone ID VFC138 (Link to dict...yBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U15456-1 Original site URL http://dict...ducing significant alignments: (bits) Value N Y16962 |Y16962.1 Dictyostelium discoideum mRNA for cathepsin D.... 815 0.0 4 AJ243946 |AJ243946.1 Dictyostelium discoideum ctsD gene for cathepsin D, exons 1 to 2. 815 0.0 5

TgrC1 mediates cell-cell adhesion by interacting with TgrB1 via mutual IPT/TIG domains during development of Dictyostelium discoideum.

Science.gov (United States)

Chen, Gong; Wang, Jun; Xu, Xiaoqun; Wu, Xiangfu; Piao, Ruihan; Siu, Chi-Hung

2013-06-01

Cell-cell adhesion plays crucial roles in cell differentiation and morphogenesis during development of Dictyostelium discoideum. The heterophilic adhesion protein TgrC1 (Tgr is transmembrane, IPT, IG, E-set, repeat protein) is expressed during cell aggregation, and disruption of the tgrC1 gene results in the arrest of development at the loose aggregate stage. We have used far-Western blotting coupled with MS to identify TgrB1 as the heterophilic binding partner of TgrC1. Co-immunoprecipitation and pull-down studies showed that TgrB1 and TgrC1 are capable of binding with each other in solution. TgrB1 and TgrC1 are encoded by a pair of adjacent genes which share a common promoter. Both TgrB1 and TgrC1 are type I transmembrane proteins, which contain three extracellular IPT/TIG (immunoglobulin, plexin, transcription factor-like/transcription factor immunoglobulin) domains. Antibodies raised against TgrB1 inhibit cell reassociation at the post-aggregation stage of development and block fruiting body formation. Ectopic expression of TgrB1 and TgrC1 driven by the actin15 promoter leads to heterotypic cell aggregation of vegetative cells. Using recombinant proteins that cover different portions of TgrB1 and TgrC1 in binding assays, we have mapped the cell-binding regions in these two proteins to Lys(537)-Ala(783) in TgrB1 and Ile(336)-Val(360) in TgrC1, corresponding to their respective TIG3 and TIG2 domain.

Dicty_cDB: AFL289 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available AF (Link to library) AFL289 (Link to dictyBase) - - - - AFL289F (Link to Original s...ite) AFL289F 123 - - - - - - Show AFL289 Library AF (Link to library) Clone ID AFL289 (Link to dictyBase) Atlas ID - NBRP ID - dict...yBase ID - Link to Contig - Original site URL http://dictycdb.biol.tsukuba.ac.jp/CSM/... E Sequences producing significant alignments: (bits) Value N X51892 |X51892.1 Dict...yostelium discoideum SP60 gene for spore coat protein. 82 5e-29 2 X52105 |X52105.1 Dictyostelium discoideu

Predicting the distribution of spiral waves from cell properties in a developmental-path model of Dictyostelium pattern formation.

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Daniel Geberth

2009-07-01

Full Text Available The slime mold Dictyostelium discoideum is one of the model systems of biological pattern formation. One of the most successful answers to the challenge of establishing a spiral wave pattern in a colony of homogeneously distributed D. discoideum cells has been the suggestion of a developmental path the cells follow (Lauzeral and coworkers. This is a well-defined change in properties each cell undergoes on a longer time scale than the typical dynamics of the cell. Here we show that this concept leads to an inhomogeneous and systematic spatial distribution of spiral waves, which can be predicted from the distribution of cells on the developmental path. We propose specific experiments for checking whether such systematics are also found in data and thus, indirectly, provide evidence of a developmental path.

Dicty_cDB: [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VF (Link to library) VFB752 (Link to dictyBase) - - - Contig-U14717-1 VFB752E (Link...) Clone ID VFB752 (Link to dictyBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U14717-1 Ori...s: (bits) Value N AC116984 |AC116984.2 Dictyostelium discoideum chromosome 2 map 2567470-3108875 strain AX4,... complete sequence. 1215 0.0 11 AC115594 |AC115594.2 Dictyostelium discoideum chr...omosome 2 map 4071862-4101005 strain AX4, complete sequence. 113 3e-47 8 AC116920 |AC116920.2 Dictyostelium

Dicty_cDB: CFC194 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available equences producing significant alignments: (bits) Value N AF305060 |AF305060.1 Dictyostelium discoideum Wiscott...nts: (bits) Value AF305060_1( AF305060 |pid:none) Dictyostelium discoideum Wiscott

Dicty_cDB: AFE360 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available t alignments: (bits) Value N AF305060 |AF305060.1 Dictyostelium discoideum Wiscott-Aldrich syndrome protein ...(bits) Value AF305060_1( AF305060 |pid:none) Dictyostelium discoideum Wiscott-A..

Phg1/TM9 proteins control intracellular killing of bacteria by determining cellular levels of the Kil1 sulfotransferase in Dictyostelium.

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Marion Le Coadic

Full Text Available Dictyostelium discoideum has largely been used to study phagocytosis and intracellular killing of bacteria. Previous studies have shown that Phg1A, Kil1 and Kil2 proteins are necessary for efficient intracellular killing of Klebsiella bacteria. Here we show that in phg1a KO cells, cellular levels of lysosomal glycosidases and lysozyme are decreased, and lysosomal pH is increased. Surprisingly, overexpression of Kil1 restores efficient killing in phg1a KO cells without correcting these lysosomal anomalies. Conversely, kil1 KO cells are defective for killing, but their enzymatic content and lysosomal pH are indistinguishable from WT cells. The killing defect of phg1a KO cells can be accounted for by the observation that in these cells the stability and the cellular amount of Kil1 are markedly reduced. Since Kil1 is the only sulfotransferase characterized in Dictyostelium, an (unidentified sulfated factor, defective in both phg1a and kil1 KO cells, may play a key role in intracellular killing of Klebsiella bacteria. In addition, Phg1B plays a redundant role with Phg1A in controlling cellular amounts of Kil1 and intracellular killing. Finally, cellular levels of Kil1 are unaffected in kil2 KO cells, and Kil1 overexpression does not correct the killing defect of kil2 KO cells, suggesting that Kil2 plays a distinct role in intracellular killing.

The ROCO Kinase QkgA Is Necessary for Proliferation Inhibition by Autocrine Signals in Dictyostelium discoideum▿

OpenAIRE

Phillips, Jonathan E.; Gomer, Richard H.

2010-01-01

AprA and CfaD are secreted proteins that function as autocrine signals to inhibit cell proliferation in Dictyostelium discoideum. Cells lacking AprA or CfaD proliferate rapidly, and adding AprA or CfaD to cells slows proliferation. Cells lacking the ROCO kinase QkgA proliferate rapidly, with a doubling time 83% of that of the wild type, and overexpression of a QkgA-green fluorescent protein (GFP) fusion protein slows cell proliferation. We found that qkgA− cells accumulate normal levels of ex...

dictyExpress: a web-based platform for sequence data management and analytics in Dictyostelium and beyond.

Science.gov (United States)

Stajdohar, Miha; Rosengarten, Rafael D; Kokosar, Janez; Jeran, Luka; Blenkus, Domen; Shaulsky, Gad; Zupan, Blaz

2017-06-02

Dictyostelium discoideum, a soil-dwelling social amoeba, is a model for the study of numerous biological processes. Research in the field has benefited mightily from the adoption of next-generation sequencing for genomics and transcriptomics. Dictyostelium biologists now face the widespread challenges of analyzing and exploring high dimensional data sets to generate hypotheses and discovering novel insights. We present dictyExpress (2.0), a web application designed for exploratory analysis of gene expression data, as well as data from related experiments such as Chromatin Immunoprecipitation sequencing (ChIP-Seq). The application features visualization modules that include time course expression profiles, clustering, gene ontology enrichment analysis, differential expression analysis and comparison of experiments. All visualizations are interactive and interconnected, such that the selection of genes in one module propagates instantly to visualizations in other modules. dictyExpress currently stores the data from over 800 Dictyostelium experiments and is embedded within a general-purpose software framework for management of next-generation sequencing data. dictyExpress allows users to explore their data in a broader context by reciprocal linking with dictyBase-a repository of Dictyostelium genomic data. In addition, we introduce a companion application called GenBoard, an intuitive graphic user interface for data management and bioinformatics analysis. dictyExpress and GenBoard enable broad adoption of next generation sequencing based inquiries by the Dictyostelium research community. Labs without the means to undertake deep sequencing projects can mine the data available to the public. The entire information flow, from raw sequence data to hypothesis testing, can be accomplished in an efficient workspace. The software framework is generalizable and represents a useful approach for any research community. To encourage more wide usage, the backend is open

Dicty_cDB: AFF743 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available Sequences producing significant alignments: (bits) Value N AF305060 |AF305060.1 Dictyostelium discoideum Wiscott...oducing significant alignments: (bits) Value AF305060_1( AF305060 |pid:none) Dictyostelium discoideum Wiscott

An unusual protein kinase phosphorylates the chemotactic receptor of Dictystelium discoideum

International Nuclear Information System (INIS)

Meier, K.; Klein, C.

1988-01-01

The authors report the cAMP-dependent phosphorylation of the chemotactic receptor of Dictyostelium discoideum in partially purified plasma membranes. The protein kinase responsible for receptor phosphorylation is associated with this fraction and preferentially phosphorylates the ligand-occupied form of the receptor. 8-Azido[ 32 P]cAMP labeling of the cell surface has shown that the cAMP receptor exists in two forms. A 45-kDa protein is predominant on unstimulated cells. cAMP stimulation results in an increased receptor phosphorylation such that the receptor migrates on NaDodSO 4 /PAGE as a 47-kDa protein. Phosphorylation of the chemotactic receptor is not detected in membrane preparations unless cAMP is added to the incubation mixture. Only under those conditions is the phosphorylated 47-kDa form observed. The requirement for cAMP reflects the fact that the kinase involved preferentially uses the ligand-occupied receptor as a substrate. In vitro phosphorylation of the receptor does not involve tyrosine residues. The enzyme does not appear to be a cAMP- or cGMP-dependent protein kinase nor is it sensitive to guanine nucleotides, Ca 2+ /calmodulin, Ca 2+ /phospholipid, or EGTA. Similarities with the β-adrenergic receptor protein kinase are discussed

Dicty_cDB: SFC807 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available icant alignments: (bits) Value N AF305060 |AF305060.1 Dictyostelium discoideum Wiscott-Aldrich syndrome prot...ng significant alignments: (bits) Value AF305060_1( AF305060 |pid:none) Dictyostelium discoideum Wiscott

Two distinct sensing pathways allow recognition of Klebsiella pneumoniae by Dictyostelium amoebae.

Science.gov (United States)

Lima, Wanessa C; Balestrino, Damien; Forestier, Christiane; Cosson, Pierre

2014-03-01

Recognition of bacteria by metazoans is mediated by receptors that recognize different types of microorganisms and elicit specific cellular responses. The soil amoebae Dictyostelium discoideum feeds upon a variable mixture of environmental bacteria, and it is expected to recognize and adapt to various food sources. To date, however, no bacteria-sensing mechanisms have been described. In this study, we isolated a Dictyostelium mutant (fspA KO) unable to grow in the presence of non-capsulated Klebsiella pneumoniae bacteria, but growing as efficiently as wild-type cells in the presence of other bacteria, such as Bacillus subtilis. fspA KO cells were also unable to respond to K. pneumoniae and more specifically to bacterially secreted folate in a chemokinetic assay, while they responded readily to B. subtilis. Remarkably, both WT and fspA KO cells were able to grow in the presence of capsulated LM21 K. pneumoniae, and responded to purified capsule, indicating that capsule recognition may represent an alternative, FspA-independent mechanism for K. pneumoniae sensing. When LM21 capsule synthesis genes were deleted, growth and chemokinetic response were lost for fspA KO cells, but not for WT cells. Altogether, these results indicate that Dictyostelium amoebae use specific recognition mechanisms to respond to different K. pneumoniae elements. © 2013 John Wiley & Sons Ltd.

Dicty_cDB: VFN644 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available A clone:ddv28g12, 3' ... 404 0.0 5 ( AF305060 ) Dictyostelium discoideum Wiscott-...ng significant alignments: (bits) Value AF305060_1( AF305060 |pid:none) Dictyostelium discoideum Wiscott-A..

Dicty_cDB: AFG687 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available ant alignments: (bits) Value N AF305060 |AF305060.1 Dictyostelium discoideum Wiscott...n Score E Sequences producing significant alignments: (bits) Value AF305060_1( AF305060 |pid:none) Dictyostelium discoideum Wiscott

NCBI nr-aa BLAST: CBRC-DDIS-01-0111 [SEVENS

Lifescience Database Archive (English)

Full Text Available CBRC-DDIS-01-0111 ref|XP_646081.1| alkaline dihydroceramidase [Dictyostelium discoi...deum AX4] gb|EAL72137.1| alkaline dihydroceramidase [Dictyostelium discoideum AX4] XP_646081.1 1e-164 100% ...

Dicty_cDB: [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VF (Link to library) VFB733 (Link to dictyBase) - - - Contig-U16279-1 VFB733P (Link... to Original site) VFB733F 608 VFB733Z 663 VFB733P 1271 - - Show VFB733 Library VF (Link to library) Clone ID VFB733 (Link to dict...yBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U16279-1 Original site URL http://dict...es producing significant alignments: (bits) Value N U23957 |U23957.1 Dictyostelium discoideum P52D mRNA, com...15 e-114 U23957_1( U23957 |pid:none) Dictyostelium discoideum P52D mRNA, co... 41

Dicty_cDB: VFA863 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available ore E Sequences producing significant alignments: (bits) Value N AF305060 |AF305060.1 Dictyostelium discoideum Wiscott...gnificant alignments: (bits) Value AF305060_1( AF305060 |pid:none) Dictyostelium discoideum Wiscott-A... 83

Dicty_cDB: CFG253 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available Value N AF305060 |AF305060.1 Dictyostelium discoideum Wiscott-Aldrich syndrome protein (wasA) gene, complete...ences producing significant alignments: (bits) Value AF305060_1( AF305060 |pid:none) Dictyostelium discoideum Wiscott

Variation in the excitability of developed D. discoideum cells as a function of agar concentration in the substrate

Science.gov (United States)

Oikawa, Noriko; Bae, Albert; Amselem, Gabriel; Bodenschatz, Eberhard

2010-03-01

In the absence of nutrients, Dictyostelium discoideum cells enter a developmental cycle--they signal each other, aggregate, and ultimately form fruiting bodies. During the signaling stage, the cells relay waves of cyclic adenosine 3',5' monophosphate (cAMP). We observed a transition from spiral to circular patterns in the signaling wave, depending on the agar concentration of the substrate. In this talk we will present the changes in the times for the onset of signaling and synchronization versus agar concentration, as measured by spectral entropy. We also will discuss the origin of these effects.

Dicty_cDB: SSG705 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available NA Score E Sequences producing significant alignments: (bits) Value N AF305060 |AF305060.1 Dictyostelium discoideum Wiscott...quences producing significant alignments: (bits) Value AF305060_1( AF305060 |pid:none) Dictyostelium discoideum Wiscott

Dicty_cDB: SFC789 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available ments: (bits) Value N AF305060 |AF305060.1 Dictyostelium discoideum Wiscott-Aldri...ne) Dictyostelium discoideum Wiscott-A... 34 0.031 protein update 2009. 1. 7 PSORT psg: 0.84 gvh: 0.42 alm:

Dicty_cDB: AFL826 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available AF (Link to library) AFL826 (Link to dictyBase) - - - - AFL826P (Link to Original s...ite) AFL826F 588 AFL826Z 766 AFL826P 1334 - - Show AFL826 Library AF (Link to library) Clone ID AFL826 (Link to dict...yBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig - Original site URL http://dictycdb.biol.t... Score E Sequences producing significant alignments: (bits) Value N ( BJ346543 ) Dictyostelium discoideum cD...NA clone:dda24d08, 3' ... 1100 0.0 1 ( BJ341682 ) Dictyostelium discoideum cDNA c

Nucleus-associated phosphorylation of Ins(1,4,5)P3 to InsP6 in Dictyostelium

NARCIS (Netherlands)

Kaay, Jeroen van der; Wesseling, Jelle; Haastert, Peter J.M. van

1995-01-01

Although many cells contain large amounts of InsP(6), its metabolism and function is still largely unknown. In Dictyostelium lysates, the formation of InsP(6) by sequential phosphorylation of inositol via Ins(3,4,6)P-3 has been described [Stevens and Irvine (1990) Nature (London) 346, 580-583]; the

Burkholderia bacteria infectiously induce the proto-farming symbiosis of Dictyostelium amoebae and food bacteria.

Science.gov (United States)

DiSalvo, Susanne; Haselkorn, Tamara S; Bashir, Usman; Jimenez, Daniela; Brock, Debra A; Queller, David C; Strassmann, Joan E

2015-09-08

Symbiotic associations can allow an organism to acquire novel traits by accessing the genetic repertoire of its partner. In the Dictyostelium discoideum farming symbiosis, certain amoebas (termed "farmers") stably associate with bacterial partners. Farmers can suffer a reproductive cost but also gain beneficial capabilities, such as carriage of bacterial food (proto-farming) and defense against competitors. Farming status previously has been attributed to amoeba genotype, but the role of bacterial partners in its induction has not been examined. Here, we explore the role of bacterial associates in the initiation, maintenance, and phenotypic effects of the farming symbiosis. We demonstrate that two clades of farmer-associated Burkholderia isolates colonize D. discoideum nonfarmers and infectiously endow them with farmer-like characteristics, indicating that Burkholderia symbionts are a major driver of the farming phenomenon. Under food-rich conditions, Burkholderia-colonized amoebas produce fewer spores than uncolonized counterparts, with the severity of this reduction being dependent on the Burkholderia colonizer. However, the induction of food carriage by Burkholderia colonization may be considered a conditionally adaptive trait because it can confer an advantage to the amoeba host when grown in food-limiting conditions. We observed Burkholderia inside and outside colonized D. discoideum spores after fruiting body formation; this observation, together with the ability of Burkholderia to colonize new amoebas, suggests a mixed mode of symbiont transmission. These results change our understanding of the D. discoideum farming symbiosis by establishing that the bacterial partner, Burkholderia, is an important causative agent of the farming phenomenon.

Dicty_cDB: SLH341 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available 5 ( AF305060 ) Dictyostelium discoideum Wiscott-Aldrich syndrome... 404 0.0 5 ( B...ts) Value AF305060_1( AF305060 |pid:none) Dictyostelium discoideum Wiscott-A... 83 8e-15 AC117076_18( AC1170

Gene discovery by chemical mutagenesis and whole-genome sequencing in Dictyostelium.

Science.gov (United States)

Li, Cheng-Lin Frank; Santhanam, Balaji; Webb, Amanda Nicole; Zupan, Blaž; Shaulsky, Gad

2016-09-01

Whole-genome sequencing is a useful approach for identification of chemical-induced lesions, but previous applications involved tedious genetic mapping to pinpoint the causative mutations. We propose that saturation mutagenesis under low mutagenic loads, followed by whole-genome sequencing, should allow direct implication of genes by identifying multiple independent alleles of each relevant gene. We tested the hypothesis by performing three genetic screens with chemical mutagenesis in the social soil amoeba Dictyostelium discoideum Through genome sequencing, we successfully identified mutant genes with multiple alleles in near-saturation screens, including resistance to intense illumination and strong suppressors of defects in an allorecognition pathway. We tested the causality of the mutations by comparison to published data and by direct complementation tests, finding both dominant and recessive causative mutations. Therefore, our strategy provides a cost- and time-efficient approach to gene discovery by integrating chemical mutagenesis and whole-genome sequencing. The method should be applicable to many microbial systems, and it is expected to revolutionize the field of functional genomics in Dictyostelium by greatly expanding the mutation spectrum relative to other common mutagenesis methods. © 2016 Li et al.; Published by Cold Spring Harbor Laboratory Press.

Nanovesicles released by Dictyostelium cells: a potential carrier for drug delivery.

Science.gov (United States)

Lavialle, Françoise; Deshayes, Sophie; Gonnet, Florence; Larquet, Eric; Kruglik, Sergei G; Boisset, Nicolas; Daniel, Régis; Alfsen, Annette; Tatischeff, Irène

2009-10-01

Nanovesicles released by Dictyostelium discoideum cells grown in the presence of the DNA-specific dye Hoechst 33342 have been previously shown to mediate the transfer of the dye into the nuclei of Hoechst-resistant cells. The present investigation extends this work by conducting experiments in the presence of hypericin, a fluorescent therapeutic photosensitizer assayed for antitumoral photodynamic therapy. Nanovesicles released by Dictyostelium cells exhibit an averaged diameter between 50 and 150 nm, as measured by transmission cryoelectron microscopy. A proteomic analysis reveals a predominance of actin and actin-related proteins. The detection of a lysosomal membrane protein (LIMP II) indicates that these vesicles are likely generated in the late endosomal compartment. The use of the hypericin-containing nanovesicles as nanodevices for in vitro drug delivery was investigated by fluorescence microscopy. The observed signal was almost exclusively located in the perinuclear area of two human cell lines, skin fibroblasts (HS68) and cervix carcinoma (HeLa) cells. Studies by confocal microscopy with specific markers of cell organelles, provided evidence that hypericin was accumulated in the Golgi apparatus. All these data shed a new light on in vitro drug delivery by using cell-released vesicles as carriers.

Dicty_cDB: SHE721 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available E Sequences producing significant alignments: (bits) Value N AF305060 |AF305060.1 Dictyostelium discoideum Wiscott...305060_1( AF305060 |pid:none) Dictyostelium discoideum Wiscott-A... 83 8e-15 AC117076_18( AC117076 |pid:none

The PsB glycoprotein complex is secreted as a preassembled precursor of the spore coat in Dictyostelium discoideum.

Science.gov (United States)

Watson, N; McGuire, V; Alexander, S

1994-09-01

The PsB glycoprotein in Dictyostelium discoideum is one of a diverse group of developmentally regulated, prespore-cell-specific proteins, that contain a common O-linked oligosaccharide. This post-translational modification is dependent on the wild-type modB allele. The PsB protein exists as part of a multiprotein complex of six different proteins, which have different post-translational modifications and are held together by both covalent and non-covalent interactions (Watson et al. (1993). J. Biol. Chem. 268, 22634-22641). In this study we have used microscopic and biochemical analyses to examine the cellular localization and function of the PsB complex during development. We found that the PsB complex first accumulates in prespore vesicles in slug cells and is secreted later during culmination and becomes localized to both the extracellular matrix of the apical spore mass of mature fruiting bodies and to the inner layer of the spore coat. The PsB associated with the spore coat is covalently bound by disulfide bridges. The PsB protein always exists in a multiprotein complex, but the composition of the PsB complex changes during secretion and spore maturation. Some of the PsB complex proteins have been identified as spore coat proteins. These data demonstrate that some of the proteins that form the spore coat exist as a preassembled precursor complex. The PsB complex is secreted in a developmentally regulated manner during the process of spore differentiation, at which time proteins of the complex, as well as additional spore coat proteins, become covalently associated in at least two forms of extracellular matrix: the interspore matrix and the spore coat. These and other studies show that proteins with modB dependent O-linked oligosaccharides are involved in a wide variety of processes underlying morphogenesis in this organism. These developmental processes are the direct result of cellular mechanisms regulating protein targeting, assembly and secretion, and the

Dicty_cDB: VSC304 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available 05060 |AF305060.1 Dictyostelium discoideum Wiscott-Aldrich syndrome protein (wasA) gene, complete cds. 622 0...bits) Value AF305060_1( AF305060 |pid:none) Dictyostelium discoideum Wiscott-A... 167 1e-44 AC117076_18( AC1

Dicty_cDB: CFG349 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available its) Value N AF305060 |AF305060.1 Dictyostelium discoideum Wiscott-Aldrich syndrome protein (wasA) gene, com...060_1( AF305060 |pid:none) Dictyostelium discoideum Wiscott-A... 247 4e-64 DQ985464_1( DQ985464 |pid:none) S

Dicty_cDB: SFG565 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available 5060.1 Dictyostelium discoideum Wiscott-Aldrich syndrome protein (wasA) gene, complete cds. 626 0.0 8 AC1170...bits) Value AF305060_1( AF305060 |pid:none) Dictyostelium discoideum Wiscott-A... 254 4e-77 AC117076_18( AC1

NCBI nr-aa BLAST: CBRC-DDIS-03-0190 [SEVENS

Lifescience Database Archive (English)

Full Text Available CBRC-DDIS-03-0190 ref|XP_646956.1| ARID/BRIGHT DNA binding domain-containing protei...n [Dictyostelium discoideum AX4] gb|EAL72978.1| ARID/BRIGHT DNA binding domain-containing protein [Dictyostelium discoideum AX4] XP_646956.1 1e-06 23% ...

Dicty_cDB: SSC474 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available SS (Link to library) SSC474 (Link to dictyBase) - - - Contig-U07719-1 SSC474P (Link... to Original site) SSC474F 368 SSC474Z 238 SSC474P 606 - - Show SSC474 Library SS (Link to library) Clone ID SSC474 (Link to dict...yBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U07719-1 Original site URL http://dict...logy vs DNA Score E Sequences producing significant alignments: (bits) Value N ( AU071762 ) Dictyostelium di...scoideum slug cDNA, clone SSC474. 448 e-121 1 ( AU060185 ) Dictyostelium discoideum slug cDNA, clone SLA535.

Dicty_cDB: [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VF (Link to library) VFB862 (Link to dictyBase) - - - Contig-U16311-1 VFB862P (Link... to Original site) VFB862F 624 VFB862Z 720 VFB862P 1344 - - Show VFB862 Library VF (Link to library) Clone ID VFB862 (Link to dict...yBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U16311-1 Original site URL http://dict...s) Value N U72746 |U72746.1 Dictyostelium discoideum cysteine proteinase (cprG) m...RNA, complete cds. 1209 0.0 5 U72745 |U72745.1 Dictyostelium discoideum cysteine proteinase (cprF) mRNA, com

Dicty_cDB: [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VF (Link to library) VFB769 (Link to dictyBase) - - - Contig-U15456-1 VFB769P (Link... to Original site) VFB769F 625 VFB769Z 690 VFB769P 1315 - - Show VFB769 Library VF (Link to library) Clone ID VFB769 (Link to dict...yBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U15456-1 Original site URL http://dict...roducing significant alignments: (bits) Value N Y16962 |Y16962.1 Dictyostelium discoideum mRNA for cathepsin... D. 1257 0.0 3 AJ243946 |AJ243946.1 Dictyostelium discoideum ctsD gene for cathep

Effect of drugs on lipid methylation, receptor-adenylate cyclase coupling and cyclic AMP secretion in Dictyostelium discoideum

NARCIS (Netherlands)

Van Waarde, Aren; Van Haastert, P.J.M.

1986-01-01

Intercellular communication in Dictyostelium discoldeum takes place by means of cyclic AMP-induced cyclic AMP-synthesis and secretion. Since phospholipid methylation has been suggested to play a role in receptor-adenylate cyclase coupling, we examined the effects of transmethylation inhibitors on

Dicty_cDB: Contig-U16102-1 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available 0 6 ( BJ408668 ) Dictyostelium discoideum cDNA clone:dds46g14, 3' ... 44 3.0 2 ( CV162186 ) CS_hyp_01d11_M13Reverse Blue crab hypoder...08_M13Reverse Blue crab hypodermis, nor... 42 3.6 2 ( AM474408 ) Vitis vinifera contig VV78X173370.5, whole

Dicty_cDB: FC-AM13 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available in (NS35) gene, complete cds. 52 0.006 1 AB022770 |AB022770.1 Human rotavirus mRNA for NSP2, complete cds. 5...9.2 Dictyostelium discoideum chromosome 2 map 4229098-4354721 strain AX4, complete sequence. 34 0.004 7 X57944 |X57944.1 Human rotavi...rus segment 7 gp33 gene. 52 0.006 1 L04534 |L04534.1 Rotavirus non-structural prote

Comparing the Dictyostelium and Entamoeba genomes reveals an ancient split in the Conosa lineage.

Directory of Open Access Journals (Sweden)

Jie Song

2005-12-01

Full Text Available The Amoebozoa are a sister clade to the fungi and the animals, but are poorly sampled for completely sequenced genomes. The social amoeba Dictyostelium discoideum and amitochondriate pathogen Entamoeba histolytica are the first Amoebozoa with genomes completely sequenced. Both organisms are classified under the Conosa subphylum. To identify Amoebozoa-specific genomic elements, we compared these two genomes to each other and to other eukaryotic genomes. An expanded phylogenetic tree built from the complete predicted proteomes of 23 eukaryotes places the two amoebae in the same lineage, although the divergence is estimated to be greater than that between animals and fungi, and probably happened shortly after the Amoebozoa split from the opisthokont lineage. Most of the 1,500 orthologous gene families shared between the two amoebae are also shared with plant, animal, and fungal genomes. We found that only 42 gene families are distinct to the amoeba lineage; among these are a large number of proteins that contain repeats of the FNIP domain, and a putative transcription factor essential for proper cell type differentiation in D. discoideum. These Amoebozoa-specific genes may be useful in the design of novel diagnostics and therapies for amoebal pathologies.

Dicty_cDB: SSC836 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available SS (Link to library) SSC836 (Link to dictyBase) - - - - SSC836E (Link to Original s...ite) - - - - - - SSC836E 502 Show SSC836 Library SS (Link to library) Clone ID SSC836 (Link to dictyBase) Atlas ID - NBRP ID - dict...yBase ID - Link to Contig - Original site URL http://dictycdb.biol.tsukuba.ac.jp/CSM/...t alignments: (bits) Value N M77492 |M77492.1 Dictyostelium discoideum glycoprotein phosphorylase 2 (glpD) g...ene, complete cds. 779 0.0 1 AC116984 |AC116984.2 Dictyostelium discoideum chromo

Dicty_cDB: [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VF (Link to library) VFB614 (Link to dictyBase) - - - Contig-U16382-1 VFB614Z (Link... to Original site) - - VFB614Z 219 - - - - Show VFB614 Library VF (Link to library) Clone ID VFB614 (Link to dict...yBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U16382-1 Original site URL http://dict...ments: (bits) Value N AC116957 |AC116957.2 Dictyostelium discoideum chromosome 2 ...tin mRNA ITL-1, 3' end. 339 9e-90 1 AC116986 |AC116986.2 Dictyostelium discoideum chromosome 2 map 2234041-2

Mechano-chemical signaling maintains the rapid movement of Dictyostelium cells

International Nuclear Information System (INIS)

Lombardi, M.L.; Knecht, D.A.; Lee, J.

2008-01-01

The survival of Dictyostelium cells depends on their ability to efficiently chemotax, either towards food or to form multicellular aggregates. Although the involvement of Ca 2+ signaling during chemotaxis is well known, it is not clear how this regulates cell movement. Previously, fish epithelial keratocytes have been shown to display transient increases in intracellular calcium ([Ca 2+ ] i ) that are mediated by stretch-activated calcium channels (SACs), which play a role in retraction of the cell body [J. Lee, A. Ishihara, G. Oxford, B. Johnson, and K. Jacobson, Regulation of cell movement is mediated by stretch-activated calcium channels. Nature, 1999. 400(6742): p. 382-6.]. To investigate the involvement of SACs in Dictyostelium movement we performed high resolution calcium imaging in wild-type (NC4A2) Dictyostelium cells to detect changes in [Ca 2+ ] i . We observed small, brief, Ca 2+ transients in randomly moving wild-type cells that were dependent on both intracellular and extracellular sources of calcium. Treatment of cells with the SAC blocker gadolinium (Gd 3+ ) inhibited transients and decreased cell speed, consistent with the involvement of SACs in regulating Dictyostelium motility. Additional support for SAC activity was given by the increase in frequency of Ca 2+ transients when Dictyostelium cells were moving on a more adhesive substratum or when they were mechanically stretched. We conclude that mechano-chemical signaling via SACs plays a major role in maintaining the rapid movement of Dictyostelium cells

Deletion of Dictyostelium discoideum Sir2A impairs cell proliferation ...

Indian Academy of Sciences (India)

Rakhee Lohia

2018-04-13

Apr 13, 2018 ... Multicellular structures developed were collected, fixed and stained with X-gal as ... classification for Sirtuins (Fyre 2000), the above were clas- ... RNA, protein and lipid substrates (Klug 1999; Hall 2005;. Gamsjaeger et al.

Dicty_cDB: CFH668 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available CF (Link to library) CFH668 (Link to dictyBase) - - - Contig-U13860-1 - (Link to Or...iginal site) - - CFH668Z 651 - - - - Show CFH668 Library CF (Link to library) Clone ID CFH668 (Link to dicty...Base) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U13860-1 Original site URL http://dictycdb.b...s) Value N AC116987 |AC116987.2 Dictyostelium discoideum chromosome 2 map 3527441-3568052 strain AX4, comple...te sequence. 58 1e-11 6 AC116305 |AC116305.2 Dictyostelium discoideum chromosome

Dicty_cDB: VHK202 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VH (Link to library) VHK202 (Link to dictyBase) - - - Contig-U16260-1 VHK202P (Link to Original site) VHK2...02F 618 VHK202Z 741 VHK202P 1339 - - Show VHK202 Library VH (Link to library) Clone ID VHK2...e URL http://dictycdb.biol.tsukuba.ac.jp/CSM/VH/VHK2-A/VHK202Q.Seq.d/ Representative seq. ID VHK2...02P (Link to Original site) Representative DNA sequence >VHK202 (VHK202Q) /CSM/VH/VHK2-A/VHK2...446805 ) Dictyostelium discoideum cDNA clone:ddv63k21, 3' ... 1449 0.0 1 ( BJ446732 ) Dictyostelium discoide

Dicty_cDB: VHK254 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VH (Link to library) VHK254 (Link to dictyBase) - - - Contig-U16260-1 VHK254P (Link to Original site) VHK2...54F 616 VHK254Z 744 VHK254P 1340 - - Show VHK254 Library VH (Link to library) Clone ID VHK2...e URL http://dictycdb.biol.tsukuba.ac.jp/CSM/VH/VHK2-C/VHK254Q.Seq.d/ Representative seq. ID VHK2...54P (Link to Original site) Representative DNA sequence >VHK254 (VHK254Q) /CSM/VH/VHK2-C/VHK2...BJ446805 ) Dictyostelium discoideum cDNA clone:ddv63k21, 3' ... 1455 0.0 1 ( BJ446732 ) Dictyostelium discoi

Dicty_cDB: SLH678 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available SL (Link to library) SLH678 (Link to dictyBase) - - - Contig-U04159-1 SLH678E (Link... to Original site) - - - - - - SLH678E 373 Show SLH678 Library SL (Link to library) Clone ID SLH678 (Link to dict...yBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U04159-1 Original site URL http://dict... producing significant alignments: (bits) Value N ( AU039970 ) Dictyostelium discoideum slug cDNA, clone SLG...865. 323 e-129 2 ( AU039381 ) Dictyostelium discoideum slug cDNA, clone SLH678. 3

Dicty_cDB: SSB373 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available SS (Link to library) SSB373 (Link to dictyBase) - G00609 DDB0216216 Contig-U04543-1...nk to library) Clone ID SSB373 (Link to dictyBase) Atlas ID - NBRP ID G00609 dictyBase ID DDB0216216 Link to... Contig Contig-U04543-1 Original site URL http://dictycdb.biol.tsukuba.ac.jp/CSM/...Value N AB016728 |AB016728.1 Dictyostelium discoideum sapA mRNA for saposin A, co... 44 7e-04 12 AC116330 |AC116330.2 Dictyostelium discoideum chromosome 2 map 3191214-3323468 strain AX4, comp

Dicty_cDB: [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VF (Link to library) VFC129 (Link to dictyBase) - - - Contig-U16543-1 VFC129F (Link... to Original site) VFC129F 276 - - - - - - Show VFC129 Library VF (Link to library) Clone ID VFC129 (Link to dict...yBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U16543-1 Original site URL http://dict... vs DNA Score E Sequences producing significant alignments: (bits) Value N M91382 |M91382.1 Dictyostelium di...scoideum thioredoxin (TRX2) mRNA, 5' end. 281 2e-96 3 M91384 |M91384.1 Dictyostelium discoideum thioredoxin

Dicty_cDB: [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VF (Link to library) VFC191 (Link to dictyBase) - - - Contig-U16281-1 VFC191F (Link... to Original site) VFC191F 350 - - - - - - Show VFC191 Library VF (Link to library) Clone ID VFC191 (Link to dict...yBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U16281-1 Original site URL http://dict...ts) Value N AC116305 |AC116305.2 Dictyostelium discoideum chromosome 2 map 1005175-1418323 strain AX4, compl... 186 3e-46 AC116305_8( AC116305 |pid:none) Dictyostelium discoideum chromosom...

Dicty_cDB: [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VF (Link to library) VFB843 (Link to dictyBase) - - - Contig-U15729-1 VFB843F (Link... to Original site) VFB843F 566 - - - - - - Show VFB843 Library VF (Link to library) Clone ID VFB843 (Link to dict...yBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U15729-1 Original site URL http://dict...roducing significant alignments: (bits) Value N U64830 |U64830.1 Dictyostelium discoideum AX2 protein tyrosi...ne kinase gene, complete cds. 1114 0.0 1 U01064 |U01064.1 Dictyostelium discoideum AX2 protein tyrosine kina

Desynchronization of cells on the developmental path triggers the formation of spiral waves of cAMP during Dictyostelium aggregation.

Science.gov (United States)

Lauzeral, J; Halloy, J; Goldbeter, A

1997-08-19

Whereas it is relatively easy to account for the formation of concentric (target) waves of cAMP in the course of Dictyostelium discoideum aggregation after starvation, the origin of spiral waves remains obscure. We investigate a physiologically plausible mechanism for the spontaneous formation of spiral waves of cAMP in D. discoideum. The scenario relies on the developmental path associated with the continuous changes in the activity of enzymes such as adenylate cyclase and phosphodiesterase observed during the hours that follow starvation. These changes bring the cells successively from a nonexcitable state to an excitable state in which they relay suprathreshold cAMP pulses, and then to autonomous oscillations of cAMP, before the system returns to an excitable state. By analyzing a model for cAMP signaling based on receptor desensitization, we show that the desynchronization of cells on this developmental path triggers the formation of fully developed spirals of cAMP. Developmental paths that do not correspond to the sequence of dynamic transitions no relay-relay-oscillations-relay are less able or fail to give rise to the formation of spirals.

Dicty_cDB: VHK278 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VH (Link to library) VHK278 (Link to dictyBase) - - - Contig-U16260-1 - (Link to Original site) VHK2...78F 533 - - - - - - Show VHK278 Library VH (Link to library) Clone ID VHK278 (Link to dicty...iol.tsukuba.ac.jp/CSM/VH/VHK2-D/VHK278Q.Seq.d/ Representative seq. ID - (Link to ...Original site) Representative DNA sequence >VHK278 (VHK278Q) /CSM/VH/VHK2-D/VHK278Q.Seq.d/ AACTCTCGAGTGCAAAA...BJ427875 ) Dictyostelium discoideum cDNA clone:ddv63k24, 5' ... 997 0.0 1 ( BJ427874 ) Dictyostelium discoideum cDNA clone:ddv63k2

Control of cyclin C levels during development of Dictyostelium.

Directory of Open Access Journals (Sweden)

David M Greene

2010-05-01

Full Text Available Cdk8 and its partner cyclin C form part of the mediator complex which links the basal transcription machinery to regulatory proteins. The pair are required for correct regulation of a subset of genes and have been implicated in control of development in a number of organisms including the social amoeba Dictyostelium discoideum. When feeding, Dictyostelium amoebae are unicellular but upon starvation they aggregate to form a multicellular structure which develops into a fruiting body containing spores. Cells in which the gene encoding Cdk8 has been deleted fail to enter aggregates due to a failure of early gene expression.We have monitored the expression levels of cyclin C protein during development and find levels decrease after the multicellular mound is formed. This decrease is triggered by extracellular cAMP that, in turn, is working in part through an increase in intracellular cAMP. The loss of cyclin C is coincident with a reduction in the association of Cdk8 with a high molecular weight complex in the nucleus. Overexpression of cyclin C and Cdk8 lead to an increased rate of early development, consistent with the levels being rate limiting.Overall these results show that both cyclin C and Cdk8 are regulated during development in response to extracellular signals and the levels of these proteins are important in controlling the timing of developmental processes. These findings have important implications for the role of these proteins in controlling development, suggesting that they are targets for developmental signals to regulate gene expression.

The cellulose-binding activity of the PsB multiprotein complex is required for proper assembly of the spore coat and spore viability in Dictyostelium discoideum.

Science.gov (United States)

Srinivasan, S; Griffiths, K R; McGuire, V; Champion, A; Williams, K L; Alexander, S

2000-08-01

The terminal event of spore differentiation in the cellular slime mould Dictyostelium discoideum is the assembly of the spore coat, which surrounds the dormant amoeba and allows the organism to survive during extended periods of environmental stress. The spore coat is a polarized extracellular matrix composed of glycoproteins and cellulose. The process of spore coat formation begins by the regulated secretion of spore coat proteins from the prespore vesicles (PSVs). Four of the major spore coat proteins (SP96, PsB/SP85, SP70 and SP60) exist as a preassembled multiprotein complex within the PSVs. This complete complex has an endogenous cellulose-binding activity. Mutant strains lacking either the SP96 or SP70 proteins produce partial complexes that do not have cellulose-binding activity, while mutants lacking SP60 produce a partial complex that retains this activity. Using a combination of immunofluorescence microscopy and biochemical methods we now show that the lack of cellulose-binding activity in the SP96 and SP70 mutants results in abnormally assembled spore coats and spores with greatly reduced viability. In contrast, the SP60 mutant, in which the PsB complex retains its cellulose-binding activity, produces spores with apparently unaltered structure and viability. Thus, it is the loss of the cellulose-binding activity of the PsB complex, rather than the mere loss of individual spore coat proteins, that results in compromised spore coat structure. These results support the idea that the cellulose-binding activity associated with the complete PsB complex plays an active role in the assembly of the spore coat.

Dicty_cDB: [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VF (Link to library) VFB895 (Link to dictyBase) - - - Contig-U10164-1 VFB895P (Link... to Original site) VFB895F 578 VFB895Z 699 VFB895P 1277 - - Show VFB895 Library VF (Link to library) Clone ID VFB895 (Link to dict...yBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U10164-1 Original site URL http://dict...ore E Sequences producing significant alignments: (bits) Value N AC115604 |AC115604.2 Dictyostelium discoide...um chromosome 2 map 4354771-4414991 strain AX4, complete sequence. 42 5e-06 9 M18106 |M18106.1 Dictyostelium

Dicty_cDB: SLF689 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available SL (Link to library) SLF689 (Link to dictyBase) - - - Contig-U16284-1 SLF689Z (Link... to Original site) - - SLF689Z 320 - - - - Show SLF689 Library SL (Link to library) Clone ID SLF689 (Link to dict...yBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U16284-1 Original site URL http://dict...DNA Score E Sequences producing significant alignments: (bits) Value N ( AC116305 ) Dictyostelium discoideum... chromosome 2 map 1005175... 478 e-131 1 ( AU053477 ) Dictyostelium discoideum slug cDNA, clone SLI726. 478 e-131 1 ( AU053102 ) Dict

Dicty_cDB: VHI285 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VH (Link to library) VHI285 (Link to dictyBase) - - - Contig-U16073-1 - (Link to Or...iginal site) VHI285F 136 - - - - - - Show VHI285 Library VH (Link to library) Clone ID VHI285 (Link to dicty...Base) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U16073-1 Original site URL http://dictycdb.b... DNA Score E Sequences producing significant alignments: (bits) Value N ( BJ423035 ) Dict...yostelium discoideum cDNA clone:ddv47i22, 5' ... 72 2e-27 3 ( BJ419916 ) Dictyostelium discoideum cD

Dicty_cDB: [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VF (Link to library) VFB662 (Link to dictyBase) - - - Contig-U15118-1 VFB662E (Link...) Clone ID VFB662 (Link to dictyBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U15118-1 Ori... significant alignments: (bits) Value N AF039211 |AF039211.1 Dictyostelium discoideum ADP/ATP translocase mR...NA, complete cds. 1011 0.0 2 AF100676 |AF100676.1 Dictyostelium discoideum ADP/ATP translocase gene, complet...drial 4.0 %: extracellular, including cell wall 4.0 %: vacuolar 4.0 %: vesicles of secretory system >> predict

A Dictyostelium secreted factor requires a PTEN-like phosphatase to slow proliferation and induce chemorepulsion.

Science.gov (United States)

Herlihy, Sarah E; Tang, Yitai; Gomer, Richard H

2013-01-01

In Dictyostelium discoideum, AprA and CfaD are secreted proteins that inhibit cell proliferation. We found that the proliferation of cells lacking CnrN, a phosphatase and tensin homolog (PTEN)-like phosphatase, is not inhibited by exogenous AprA and is increased by exogenous CfaD. The expression of CnrN in cnrN cells partially rescues these altered sensitivities, suggesting that CnrN is necessary for the ability of AprA and CfaD to inhibit proliferation. Cells lacking CnrN accumulate normal levels of AprA and CfaD. Like cells lacking AprA and CfaD, cnrN cells proliferate faster and reach a higher maximum cell density than wild type cells, tend to be multinucleate, accumulate normal levels of mass and protein per nucleus, and form less viable spores. When cnrN cells expressing myc-tagged CnrN are stimulated with a mixture of rAprA and rCfaD, levels of membrane-associated myc-CnrN increase. AprA also causes chemorepulsion of Dictyostelium cells, and CnrN is required for this process. Combined, these results suggest that CnrN functions in a signal transduction pathway downstream of AprA and CfaD mediating some, but not all, of the effects of AprA and CfaD.

Dicty_cDB: CHD405 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available CH (Link to library) CHD405 (Link to dictyBase) - - - Contig-U15984-1 CHD405E (Link... Clone ID CHD405 (Link to dictyBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U15984-1 Original site URL http://dict...ts) Value N AC115599 |AC115599.2 Dictyostelium discoideum chromosome 2 map 422909...8-4354721 strain AX4, complete sequence. 42 3e-11 9 AC115598 |AC115598.2 Dictyostelium discoideum chromosome... 2 map 581427-735498 strain AX4, complete sequence. 50 4e-11 11 CK417372 |CK417372.1 AUF_IpInt_56_d19 Intestine cDNA library Ict

Three-dimensional structure of a glycosylated cell surface antigen from D. discoideum: a primordial adhesion motif

International Nuclear Information System (INIS)

Mabbutt, B.C.; Swarbrick, J.; Cubeddu, L.; Hill, A.

1999-01-01

Full text: We have determined the solution structure of pre-spore specific antigen (PsA), a predominant cell surface glycoprotein from the slime mould Dictyostelium discoideum. The structure and function of this protein suggests that it serves as a molecular signal for multicellular organisation, and that it may also be an adhesion motif mediating direct cell-cell contact. PsA consists of a 90-residue N-terminal globular domain tethered to the cell membrane via a heavily O-glycosylated stalk and a GPI anchor. No homologous sequences have been identified for the N-terminal domain. At Macquarie University, the D. discoideum organism has been well developed as a eukaryotic expression host for glycosylated proteins. For NMR, we have engineered a soluble form of PsA (residues 1-122) containing the globular 'head' and the glycopeptide linker. 15 N- and 15 N/ 13 C-labelled PsA was generated in this organism via a protocol that is readily adaptable for the cost-effective production of milligram quantities of other isotopically labelled recombinant proteins. Using 3D heteronuclear NMR, we have solved the three-dimensional structure of the PsA glycoprotein. It defines an eight stranded β-sandwich of five-on-three topology in a unique arrangement. A long loop is constrained by a cis proline residue and a disulphide bond to form an opening across one end of the sandwich, exposing portions of the hydrophobic interior. We postulate that this distortion of the sandwich fold structures a binding site. Structural and dynamics information was also obtained concerning the intact glycopeptide linker of the protein, which comprises a repeating P-T-V-T motif. In our recombinant form, each Thr residue is modified by a single GlcNAc sugar. This simple structure yields interpretable NMR spectra, which show the glycosylated linker to be in extended conformation, and undergoing distinctly different mobility from the globular domain. These same sugar residues provide an ideal attachment

Dicty_cDB: [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VF (Link to library) VFC101 (Link to dictyBase) - - - Contig-U16544-1 VFC101Z (Link... to Original site) - - VFC101Z 556 - - - - Show VFC101 Library VF (Link to library) Clone ID VFC101 (Link to dict...yBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U16544-1 Original site URL http://dict...t alignments: (bits) Value N AY164994 |AY164994.1 Dictyostelium discoideum RTNLC (RTNLC) mRNA, complete cds.... 969 0.0 2 AY164656 |AY164656.1 Dictyostelium discoideum RTNLC (RTNLC) gene, complete cds. 565 0.0 3 AL71386

Dicty_cDB: [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VF (Link to library) VFB829 (Link to dictyBase) - - - - VFB829Z (Link to Original s...ite) - - VFB829Z 600 - - - - Show VFB829 Library VF (Link to library) Clone ID VFB829 (Link to dictyBase) Atlas ID - NBRP ID - dict...yBase ID - Link to Contig - Original site URL http://dictycdb.biol.tsukuba.ac.jp/CSM/... long chain poly-unsaturated fatty acids. 1189 0.0 1 AB029311 |AB029311.1 Dictyostelium discoideum Dd des5 g...ene for fatty acid desaturase, complete cds. 823 0.0 2 AB022097 |AB022097.1 Dictyostelium discoideum mRNA fo

Neuropharmacology of Purinergic Receptors in Human Submucous Plexus: Involvement of P2X1, P2X2, P2X3 Channels, P2Y and A3 Metabotropic Receptors in Neurotransmission

Science.gov (United States)

Liñán-Rico, A.; Wunderlich, JE.; Enneking, JT.; Tso, DR.; Grants, I.; Williams, KC.; Otey, A.; Michel, K.; Schemann, M.; Needleman, B.; Harzman, A.; Christofi, FL.

2015-01-01

Rationale The role of purinergic signaling in the human ENS is not well understood. We sought to further characterize the neuropharmacology of purinergic receptors in human ENS and test the hypothesis that endogenous purines are critical regulators of neurotransmission. Experimental Approach LSCM-Fluo-4-(Ca2+)-imaging of postsynaptic Ca2+ transients (PSCaTs) was used as a reporter of neural activity. Synaptic transmission was evoked by fiber tract electrical stimulation in human SMP surgical preparations. Pharmacological analysis of purinergic signaling was done in 1,556 neurons from 234 separate ganglia 107 patients; immunochemical labeling for P2XRs of neurons in ganglia from 19 patients. Real-time MSORT (Di-8-ANEPPS) imaging was used to test effects of adenosine on fast excitatory synaptic potentials (fEPSPs). Results Synaptic transmission is sensitive to pharmacological manipulations that alter accumulation of extracellular purines. Apyrase blocks PSCaTs in a majority of neurons. An ecto-NTPDase-inhibitor 6-N,N-diethyl-D-β,γ-dibromomethyleneATP or adenosine deaminase augments PSCaTs. Blockade of reuptake/deamination of eADO inhibits PSCaTs. Adenosine inhibits fEPSPs and PSCaTs (IC50=25μM), sensitive to MRS1220-antagonism (A3AR). A P2Y agonist ADPβS inhibits PSCaTs (IC50=111nM) in neurons without stimulatory ADPβS responses (EC50=960nM). ATP or a P2X1,2,2/3 (α,β-MeATP) agonist evokes fast, slow, biphasic Ca2+ transients or Ca2+ oscillations (EC50=400μM). PSCaTs are sensitive to P2X1 antagonist NF279. Low (20nM) or high (5μM) concentrations of P2X antagonist TNP-ATP block PSCaTs in different neurons; proportions of neurons with P2XR-ir follow the order P2X2>P2X1≫P2X3; P2X1+ P2X2 and P2X3+P2X2 are co-localized. RT-PCR identified mRNA-transcripts for P2X1-7,P2Y1,2,12-14R. Responsive neurons were also identified by HuC/D-ir. Conclusions Purines are critical regulators of neurotransmission in the human enteric nervous system. Purinergic signaling involves

3' : 5'-Cyclic AMP-dependent 3'

NARCIS (Netherlands)

Mato, José M.; Krens, Frans A.; Haastert, Peter J.M. van; Konijn, Theo M.

1977-01-01

Suspensions of 3':5'-cyclic AMP (cAMP)-sensitive cells of Dictyostelium discoideum responded to a cAMP pulse with increased 3':5'-cyclic GMP (cGMP) levels. Under the assay conditions used (2 × 10^8 cells per ml in 10 mM phosphate buffer, pH 6.0) cAMP (5 × 10-8 M final concentration) increased cGMP

Dicty_cDB: VFE551 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available Value N AF305060 |AF305060.1 Dictyostelium discoideum Wiscott-Aldrich syndrome protein (wasA) gene, complet...ictyostelium discoideum Wiscott-A... 112 4e-24 AC117076_18( AC117076 |pid:none) D

A Dictyostelium secreted factor requires a PTEN-like phosphatase to slow proliferation and induce chemorepulsion.

Directory of Open Access Journals (Sweden)

Sarah E Herlihy

Full Text Available In Dictyostelium discoideum, AprA and CfaD are secreted proteins that inhibit cell proliferation. We found that the proliferation of cells lacking CnrN, a phosphatase and tensin homolog (PTEN-like phosphatase, is not inhibited by exogenous AprA and is increased by exogenous CfaD. The expression of CnrN in cnrN cells partially rescues these altered sensitivities, suggesting that CnrN is necessary for the ability of AprA and CfaD to inhibit proliferation. Cells lacking CnrN accumulate normal levels of AprA and CfaD. Like cells lacking AprA and CfaD, cnrN cells proliferate faster and reach a higher maximum cell density than wild type cells, tend to be multinucleate, accumulate normal levels of mass and protein per nucleus, and form less viable spores. When cnrN cells expressing myc-tagged CnrN are stimulated with a mixture of rAprA and rCfaD, levels of membrane-associated myc-CnrN increase. AprA also causes chemorepulsion of Dictyostelium cells, and CnrN is required for this process. Combined, these results suggest that CnrN functions in a signal transduction pathway downstream of AprA and CfaD mediating some, but not all, of the effects of AprA and CfaD.

Dicty_cDB: VHK596 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available 34 4.7 AY698035_1( AY698035 |pid:none) Desmognathus marmoratus isolate 69... 33 6.1 AC116957_68( AC116957 |p...id:none) Dictyostelium discoideum chromoso... 33 8.0 AY612344_1( AY612344 |pid:none) Desmognathus marmoratus

Activation of G-proteins by receptor-stimulated nucleoside diphosphate kinase in Dictyostelium.

Science.gov (United States)

Bominaar, A A; Molijn, A C; Pestel, M; Veron, M; Van Haastert, P J

1993-01-01

Recently, interest in the enzyme nucleoside diphosphate kinase (EC2.7.4.6) has increased as a result of its possible involvement in cell proliferation and development. Since NDP kinase is one of the major sources of GTP in cells, it has been suggested that the effects of an altered NDP kinase activity on cellular processes might be the result of altered transmembrane signal transduction via guanine nucleotide-binding proteins (G-proteins). In the cellular slime mould Dictyostelium discoideum, extracellular cAMP induces an increase of phospholipase C activity via a surface cAMP receptor and G-proteins. In this paper it is demonstrated that part of the cellular NDP kinase is associated with the membrane and stimulated by cell surface cAMP receptors. The GTP produced by the action of NDP kinase is capable of activating G-proteins as monitored by altered G-protein-receptor interaction and the activation of the effector enzyme phospholipase C. Furthermore, specific monoclonal antibodies inhibit the effect of NDP kinase on G-protein activation. These results suggest that receptor-stimulated NDP kinase contributes to the mediation of hormone action by producing GTP for the activation of GTP-binding proteins. Images PMID:8389692

Dicty_cDB: CFH521 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available CF (Link to library) CFH521 (Link to dictyBase) - - - Contig-U16381-1 - (Link to Or...iginal site) CFH521F 134 - - - - - - Show CFH521 Library CF (Link to library) Clone ID CFH521 (Link to dicty...Base) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U16381-1 Original site URL http://dictycdb.b...mology vs DNA Score E Sequences producing significant alignments: (bits) Value N AC115685 |AC115685.1 Dict.... 82 2e-29 3 X51892 |X51892.1 Dictyostelium discoideum SP60 gene for spore coat protein. 82 4e-29 2 X52105 |X52105.1 Dict

The carboxy-terminal domain of Dictyostelium C-module-binding factor is an independent gene regulatory entity.

Directory of Open Access Journals (Sweden)

Jörg Lucas

Full Text Available The C-module-binding factor (CbfA is a multidomain protein that belongs to the family of jumonji-type (JmjC transcription regulators. In the social amoeba Dictyostelium discoideum, CbfA regulates gene expression during the unicellular growth phase and multicellular development. CbfA and a related D. discoideum CbfA-like protein, CbfB, share a paralogous domain arrangement that includes the JmjC domain, presumably a chromatin-remodeling activity, and two zinc finger-like (ZF motifs. On the other hand, the CbfA and CbfB proteins have completely different carboxy-terminal domains, suggesting that the plasticity of such domains may have contributed to the adaptation of the CbfA-like transcription factors to the rapid genome evolution in the dictyostelid clade. To support this hypothesis we performed DNA microarray and real-time RT-PCR measurements and found that CbfA regulates at least 160 genes during the vegetative growth of D. discoideum cells. Functional annotation of these genes revealed that CbfA predominantly controls the expression of gene products involved in housekeeping functions, such as carbohydrate, purine nucleoside/nucleotide, and amino acid metabolism. The CbfA protein displays two different mechanisms of gene regulation. The expression of one set of CbfA-dependent genes requires at least the JmjC/ZF domain of the CbfA protein and thus may depend on chromatin modulation. Regulation of the larger group of genes, however, does not depend on the entire CbfA protein and requires only the carboxy-terminal domain of CbfA (CbfA-CTD. An AT-hook motif located in CbfA-CTD, which is known to mediate DNA binding to A+T-rich sequences in vitro, contributed to CbfA-CTD-dependent gene regulatory functions in vivo.

Acanthamoeba and Dictyostelium as Cellular Models for Legionella Infection

Science.gov (United States)

Swart, A. Leoni; Harrison, Christopher F.; Eichinger, Ludwig; Steinert, Michael; Hilbi, Hubert

2018-01-01

Environmental bacteria of the genus Legionella naturally parasitize free-living amoebae. Upon inhalation of bacteria-laden aerosols, the opportunistic pathogens grow intracellularly in alveolar macrophages and can cause a life-threatening pneumonia termed Legionnaires' disease. Intracellular replication in amoebae and macrophages takes place in a unique membrane-bound compartment, the Legionella-containing vacuole (LCV). LCV formation requires the bacterial Icm/Dot type IV secretion system, which translocates literally hundreds of “effector” proteins into host cells, where they modulate crucial cellular processes for the pathogen's benefit. The mechanism of LCV formation appears to be evolutionarily conserved, and therefore, amoebae are not only ecologically significant niches for Legionella spp., but also useful cellular models for eukaryotic phagocytes. In particular, Acanthamoeba castellanii and Dictyostelium discoideum emerged over the last years as versatile and powerful models. Using genetic, biochemical and cell biological approaches, molecular interactions between amoebae and Legionella pneumophila have recently been investigated in detail with a focus on the role of phosphoinositide lipids, small and large GTPases, autophagy components and the retromer complex, as well as on bacterial effectors targeting these host factors. PMID:29552544

Acanthamoeba and Dictyostelium as Cellular Models for Legionella Infection

Directory of Open Access Journals (Sweden)

A. Leoni Swart

2018-03-01

Full Text Available Environmental bacteria of the genus Legionella naturally parasitize free-living amoebae. Upon inhalation of bacteria-laden aerosols, the opportunistic pathogens grow intracellularly in alveolar macrophages and can cause a life-threatening pneumonia termed Legionnaires' disease. Intracellular replication in amoebae and macrophages takes place in a unique membrane-bound compartment, the Legionella-containing vacuole (LCV. LCV formation requires the bacterial Icm/Dot type IV secretion system, which translocates literally hundreds of “effector” proteins into host cells, where they modulate crucial cellular processes for the pathogen's benefit. The mechanism of LCV formation appears to be evolutionarily conserved, and therefore, amoebae are not only ecologically significant niches for Legionella spp., but also useful cellular models for eukaryotic phagocytes. In particular, Acanthamoeba castellanii and Dictyostelium discoideum emerged over the last years as versatile and powerful models. Using genetic, biochemical and cell biological approaches, molecular interactions between amoebae and Legionella pneumophila have recently been investigated in detail with a focus on the role of phosphoinositide lipids, small and large GTPases, autophagy components and the retromer complex, as well as on bacterial effectors targeting these host factors.

Dicty_cDB: Contig-U16423-1 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available 65776 ) Dictyostelium discoideum cDNA clone:ddc36f16, 5' ... 74 3e-22 2 ( FG288312 ) 1108793266221 New World... 4e-19 3 ( FI057728 ) CHO_OF6610xh18r1.ab1 CHO_OF6 Nicotiana tabacum ge... 64 6e-19 3 ( FG286148 ) 1108770713996 New World...9c24,... 90 7e-18 3 ( CJ411606 ) Molgula tectiformis cDNA, larva clone:mtlv010d03,... 90 7e-18 3 ( FG288831 ) 1108793284713 New World...e-13 2 ( FG299437 ) 1108793335742 New World Screwworm Larvae 9387 EST... 80 2e-13 2 ( DV613229 ) EST1216225 ...BJ379331 ) Dictyostelium discoideum cDNA clone:ddc34c13, 3' ... 90 5e-13 1 ( FG300027 ) 1108793358668 New World

Dicty_cDB: SSL103 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available (bits) Value N AF305060 |AF305060.1 Dictyostelium discoideum Wiscott-Aldrich syndrome protein (wasA) gene, c...ctyostelium discoideum Wiscott-A... 306 4e-82 FN392319_1421( FN392319 |pid:none) Pichia pastoris GS115 chrom

Dicty_cDB: Contig-U15153-1 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available 42 ) Dictyostelium discoideum gamete cDNA, clone FC-AU21. 400 e-170 4 ( BV426376 ) S237P6125FG3.T0 Portugu...eseWaterDog Canis familiar... 52 0.020 1 ( AC068970 ) Homo sapiens BAC clone RP11-8

Dicty_cDB: SFD117 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available SF (Link to library) SFD117 (Link to dictyBase) - - - Contig-U16381-1 SFD117F (Link... to Original site) SFD117F 107 - - - - - - Show SFD117 Library SF (Link to library) Clone ID SFD117 (Link to dict...yBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U16381-1 Original site URL http://dict...s producing significant alignments: (bits) Value N AC115685 |AC115685.1 Dictyoste...lium discoideum chromosome 2 map 4718821-4752388 strain AX4, complete sequence. 80 2e-21 3 X51892 |X51892.1 Dict

Survival of the fattest

DEFF Research Database (Denmark)

Morgani, Sophie M; Brickman, Joshua M

2013-01-01

Experiments on the social amoeba Dictyostelium discoideum show that the origins of lineage bias in this system lie in the nutritional history of individual cells. Clues to the molecular basis for this process suggest similar forces may be at work in early mammalian development.......Experiments on the social amoeba Dictyostelium discoideum show that the origins of lineage bias in this system lie in the nutritional history of individual cells. Clues to the molecular basis for this process suggest similar forces may be at work in early mammalian development....

A flavin-dependent halogenase catalyzes the chlorination step in the biosynthesis of Dictyostelium differentiation-inducing factor 1.

Science.gov (United States)

Neumann, Christopher S; Walsh, Christopher T; Kay, Robert R

2010-03-30

Differentiation-inducing factor 1 (DIF-1) is a polyketide-derived morphogen which drives stalk cell formation in the developmental cycle of Dictyostelium discoideum. Previous experiments demonstrated that the biosynthetic pathway proceeds via dichlorination of the precursor molecule THPH, but the enzyme responsible for this transformation has eluded characterization. Our recent studies on prokaryotic flavin-dependent halogenases and insights from the sequenced Dd genome led us to a candidate gene for this transformation. In this work, we present in vivo and in vitro evidence that chlA from Dd encodes a flavin-dependent halogenase capable of catalyzing both chlorinations in the biosynthesis of DIF-1. The results provide in vitro characterization of a eukaryotic oxygen-dependent halogenase and demonstrate a broad reach in biology for this molecular tailoring strategy, notably its involvement in the differentiation program of a social amoeba.

Evidence for nucleolar subcompartments in Dictyostelium

International Nuclear Information System (INIS)

Catalano, Andrew; O’Day, Danton H.

2015-01-01

Highlights: • Two nucleolar subcompartments (NoSC1, NoSC2) were found in Dictyostelium. • Specific nucleolar proteins localize to different nucleolar subcompartments. • Specific proteins exit NoSC1 and NoSC2 differently upon Actinomycin D treatment. • KRKR appears to function as an NoSC2 nucleolar subcompartment localization signal. - Abstract: The nucleolus is a multifunctional nuclear compartment usually consisting of two to three subcompartments which represent stages of ribosomal biogenesis. It is linked to several human diseases including viral infections, cancer, and neurodegeneration. Dictyostelium is a model eukaryote for the study of fundamental biological processes as well as several human diseases however comparatively little is known about its nucleolus. Unlike most nucleoli it does not possess visible subcompartments at the ultrastructural level. Several recently identified nucleolar proteins in Dictyostelium leave the nucleolus after treatment with the rDNA transcription inhibitor actinomycin-D (AM-D). Different proteins exit in different ways, suggesting that previously unidentified nucleolar subcompartments may exist. The identification of nucleolar subcompartments would help to better understand the nucleolus in this model eukaryote. Here, we show that Dictyostelium nucleolar proteins nucleomorphin isoform NumA1 and Bud31 localize throughout the entire nucleolus while calcium-binding protein 4a localizes to only a portion, representing nucleolar subcompartment 1 (NoSC1). SWI/SNF complex member Snf12 localizes to a smaller area within NoSC1 representing a second nucleolar subcompartment, NoSC2. The nuclear/nucleolar localization signal KRKR from Snf12 localized GFP to NoSC2, and thus also appears to function as a nucleolar subcompartment localization signal. FhkA localizes to the nucleolar periphery displaying a similar pattern to that of Hsp32. Similarities between the redistribution patterns of Dictyostelium nucleolar proteins during

Evidence for nucleolar subcompartments in Dictyostelium

Energy Technology Data Exchange (ETDEWEB)

Catalano, Andrew, E-mail: acatalano@ccny.cuny.edu [Department of Biology, University of Toronto at Mississauga, 3359 Mississauga Rd. N., Mississauga, Ontario L5L 1C6 (Canada); O’Day, Danton H., E-mail: danton.oday@utoronto.ca [Department of Biology, University of Toronto at Mississauga, 3359 Mississauga Rd. N., Mississauga, Ontario L5L 1C6 (Canada); Department of Cell and Systems Biology, University of Toronto, 25 Harbord St., Toronto, Ontario M5S 3G5 (Canada)

2015-01-24

Highlights: • Two nucleolar subcompartments (NoSC1, NoSC2) were found in Dictyostelium. • Specific nucleolar proteins localize to different nucleolar subcompartments. • Specific proteins exit NoSC1 and NoSC2 differently upon Actinomycin D treatment. • KRKR appears to function as an NoSC2 nucleolar subcompartment localization signal. - Abstract: The nucleolus is a multifunctional nuclear compartment usually consisting of two to three subcompartments which represent stages of ribosomal biogenesis. It is linked to several human diseases including viral infections, cancer, and neurodegeneration. Dictyostelium is a model eukaryote for the study of fundamental biological processes as well as several human diseases however comparatively little is known about its nucleolus. Unlike most nucleoli it does not possess visible subcompartments at the ultrastructural level. Several recently identified nucleolar proteins in Dictyostelium leave the nucleolus after treatment with the rDNA transcription inhibitor actinomycin-D (AM-D). Different proteins exit in different ways, suggesting that previously unidentified nucleolar subcompartments may exist. The identification of nucleolar subcompartments would help to better understand the nucleolus in this model eukaryote. Here, we show that Dictyostelium nucleolar proteins nucleomorphin isoform NumA1 and Bud31 localize throughout the entire nucleolus while calcium-binding protein 4a localizes to only a portion, representing nucleolar subcompartment 1 (NoSC1). SWI/SNF complex member Snf12 localizes to a smaller area within NoSC1 representing a second nucleolar subcompartment, NoSC2. The nuclear/nucleolar localization signal KRKR from Snf12 localized GFP to NoSC2, and thus also appears to function as a nucleolar subcompartment localization signal. FhkA localizes to the nucleolar periphery displaying a similar pattern to that of Hsp32. Similarities between the redistribution patterns of Dictyostelium nucleolar proteins during

NCBI nr-aa BLAST: CBRC-DDIS-02-0043 [SEVENS

Lifescience Database Archive (English)

Full Text Available CBRC-DDIS-02-0043 gb|AAL92369.2| similar to Homo sapiens (Human). Testis intercellular mediator... Peas (Sortilin 1) (Hypothetical protein) (Testis intracellular mediator protein) [Dictyostelium discoideum] AAL92369.2 1e-178 98% ...

Deletion of P2X2 and P2X3 receptor subunits does not alter motility of the mouse colon

Directory of Open Access Journals (Sweden)

Matthew DeVries

2010-03-01

Full Text Available Purinergic P2X receptors contribute to neurotransmission in the gut. P2X receptors are ligand-gated cation channels that mediate synaptic excitation in subsets of enteric neurons. The present study evaluated colonic motility in vitro and in vivo in wild type (WT and P2X2 and P2X3 subunit knockout (KO mice. The muscarinic receptor agonist, bethanechol (0.3-3 micromolar, caused similar contractions of the longitudinal muscle in colon segments from WT, P2X2 and P2X3 subunit KO mice. Nicotine (1-300 micromolar, acting at neuronal nicotinic receptors, caused similar longitudinal muscle relaxations in colonic segments from WT and P2X2 and P2X3 subunit KO mice. Nicotine-induced relaxations were inhibited by nitro-L-arginine (NLA, 100 micromolar and apamin (0.1 micromolar which block inhibitory neuromuscular transmission. ATP (1-1000 micromolar caused contractions only in the presence of NLA and apamin. ATP-induced contractions were similar in colon segments from WT, P2X2 and P2X3 KO mice. The mouse colon generates spontaneous migrating motor complexes (MMCs in vitro. The MMC frequency was higher in P2X2 KO compared to WT tissues; other parameters of the MMC were similar in colon segments from WT, P2X2 and P2X3 KO mice. 5-Hydroxytryptophan-induced fecal output was similar in WT, P2X2 and P2X3 KO mice. These data indicate that nicotinic receptors are located predominately on inhibitory motor neurons supplying the longitudinal muscle in the mouse colon. P2X2 or P2X3 subunit containing receptors are not localized to motorneurons supplying the longitudinal muscle. Synaptic transmission mediated by P2X2 or P2X3 subunit containing receptors is not required for propulsive motility in the mouse colon.

Dicty_cDB: Contig-U04690-1 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available scauevk mixed_tissue Sebastes... 34 9.9 2 >( AU071707 ) Dictyostelium discoideum slug cDNA, clone SSC319. L... ( AP006852 ) Candida albicans genomic DNA, chromosome 7, compl... 32 9.3 2 ( GE799624 ) EST_scau_evk_888927

Dicty_cDB: VFI685 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VF (Link to library) VFI685 (Link to dictyBase) - - - Contig-U16455-1 - (Link to Or...iginal site) - - VFI685Z 189 - - - - Show VFI685 Library VF (Link to library) Clone ID VFI685 (Link to dicty...Base) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U16455-1 Original site URL http://dictycdb.b...ignments: (bits) Value N ( BJ432495 ) Dictyostelium discoideum cDNA clone:ddv18i22, 3' ... 313 1e-81 1 ( X55...973 ) D. discoideum EF1-I gene for elongation factor 1 al... 305 3e-79 1 ( AU285051 ) Dict

Dicty_cDB: [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VF (Link to library) VFB702 (Link to dictyBase) - - - Contig-U12284-1 VFB702P (Link... to Original site) VFB702F 478 VFB702Z 615 VFB702P 1093 - - Show VFB702 Library VF (Link to library) Clone ID VFB702 (Link to dict...yBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U12284-1 Original site URL http://dict... N G65021 |G65021.1 DH0281 Dictyostelium discoideum ( P.Dear ) Dictyostelium disc...8.0 %: mitochondrial 4.0 %: vacuolar 4.0 %: Golgi >> prediction for VFB702 is nuc 5' end seq. ID VFB702F 5'

Purinergní P2X rodina a specifické vlastnosti P2X7 podtypu

Czech Academy of Sciences Publication Activity Database

Jindřichová, Marie; Zemková, Hana

2013-01-01

Roč. 62, č. 2 (2013), s. 40-46 ISSN 1210-6313 R&D Projects: GA ČR(CZ) GPP304/12/P371 Institutional support: RVO:67985823 Keywords : extracellular ATP * purinergic P2X family * P2X7 receptor * cell proliferation and apoptosis Subject RIV: ED - Physiology

Dicty_cDB: CFE213 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available CF (Link to library) CFE213 (Link to dictyBase) - - - Contig-U16381-1 CFE213F (Link... to Original site) CFE213F 111 - - - - - - Show CFE213 Library CF (Link to library) Clone ID CFE213 (Link to dict...yBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U16381-1 Original site URL http://dict... E Sequences producing significant alignments: (bits) Value N AC115685 |AC115685.1 Dict...yostelium discoideum chromosome 2 map 4718821-4752388 strain AX4, complete sequence. 80 9e-24 3 X51892 |X51892.1 Dict

Dicty_cDB: CFH713 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available CF (Link to library) CFH713 (Link to dictyBase) - - - Contig-U16381-1 - (Link to Or...iginal site) CFH713F 133 - - - - - - Show CFH713 Library CF (Link to library) Clone ID CFH713 (Link to dicty...Base) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U16381-1 Original site URL http://dictycdb.b...ogy vs DNA Score E Sequences producing significant alignments: (bits) Value N X51892 |X51892.1 Dict...yostelium discoideum SP60 gene for spore coat protein. 80 2e-23 3 AC115685 |AC115685.1 Dict

Dicty_cDB: CFH518 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available CF (Link to library) CFH518 (Link to dictyBase) - - - Contig-U16381-1 - (Link to Or...iginal site) CFH518F 131 - - - - - - Show CFH518 Library CF (Link to library) Clone ID CFH518 (Link to dicty...Base) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U16381-1 Original site URL http://dictycdb.b...vs DNA Score E Sequences producing significant alignments: (bits) Value N X51892 |X51892.1 Dict...yostelium discoideum SP60 gene for spore coat protein. 80 2e-22 3 AC115685 |AC115685.1 Dictyos

Dicty_cDB: CFI225 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available CF (Link to library) CFI225 (Link to dictyBase) - - - Contig-U16381-1 - (Link to Or...iginal site) CFI225F 133 - - - - - - Show CFI225 Library CF (Link to library) Clone ID CFI225 (Link to dicty...Base) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U16381-1 Original site URL http://dictycdb.b...ogy vs DNA Score E Sequences producing significant alignments: (bits) Value N X51892 |X51892.1 Dict...yostelium discoideum SP60 gene for spore coat protein. 80 2e-23 3 AC115685 |AC115685.1 Dict

Dicty_cDB: CFH557 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available CF (Link to library) CFH557 (Link to dictyBase) - - - - - (Link to Original site) C...FH557F 132 - - - - - - Show CFH557 Library CF (Link to library) Clone ID CFH557 (Link to dictyBase) Atlas ID - NBRP ID - dict...yBase ID - Link to Contig - Original site URL http://dictycdb.biol.tsukuba.ac.jp/CSM/CF/CFH...roducing significant alignments: (bits) Value N AC115685 |AC115685.1 Dictyosteliu...m discoideum chromosome 2 map 4718821-4752388 strain AX4, complete sequence. 80 3e-28 3 X51892 |X51892.1 Dict

Dicty_cDB: [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VF (Link to library) VFC154 (Link to dictyBase) - - - Contig-U16363-1 VFC154Z (Link... to Original site) - - VFC154Z 551 - - - - Show VFC154 Library VF (Link to library) Clone ID VFC154 (Link to dict...yBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U16363-1 Original site URL http://dict...s: (bits) Value N U82513 |U82513.1 Dictyostelium discoideum random slug cDNA25 protein (rsc25) mRNA, partial...producing significant alignments: (bits) Value U82513_1( U82513 |pid:none) Dictyostelium discoideum random s

Dicty_cDB: Contig-U04404-1 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available lone:VS... 498 e-176 2 ( AU266939 ) Dictyostelium discoideum vegetative cDNA clone:VS... 68 5e-07 1 ( FG2946...24 ) 1108770716152 New World Screwworm Larvae 9387 EST... 42 0.027 2 ( CU469174 ) Pig DNA sequence *** SEQUE

Sequence analysis of RNase MRP RNA reveals its origination from eukaryotic RNase P RNA

Science.gov (United States)

Zhu, Yanglong; Stribinskis, Vilius; Ramos, Kenneth S.; Li, Yong

2006-01-01

RNase MRP is a eukaryote-specific endoribonuclease that generates RNA primers for mitochondrial DNA replication and processes precursor rRNA. RNase P is a ubiquitous endoribonuclease that cleaves precursor tRNA transcripts to produce their mature 5′ termini. We found extensive sequence homology of catalytic domains and specificity domains between their RNA subunits in many organisms. In Candida glabrata, the internal loop of helix P3 is 100% conserved between MRP and P RNAs. The helix P8 of MRP RNA from microsporidia Encephalitozoon cuniculi is identical to that of P RNA. Sequence homology can be widely spread over the whole molecule of MRP RNA and P RNA, such as those from Dictyostelium discoideum. These conserved nucleotides between the MRP and P RNAs strongly support the hypothesis that the MRP RNA is derived from the P RNA molecule in early eukaryote evolution. PMID:16540690

Dicty_cDB: [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VF (Link to library) VFC254 (Link to dictyBase) - - - Contig-U15456-1 VFC254P (Link... to Original site) VFC254F 509 VFC254Z 569 VFC254P 1078 - - Show VFC254 Library VF (Link to library) Clone ID VFC254 (Link to dict...yBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U15456-1 Original site URL http://dict...mology vs DNA Score E Sequences producing significant alignments: (bits) Value N Y16962 |Y16962.1 Dictyostel...ium discoideum mRNA for cathepsin D. 910 0.0 3 AJ243946 |AJ243946.1 Dictyostelium

Dicty_cDB: FC-IC0102 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available FC-IC (Link to library) FC-IC0102 (Link to dictyBase) - - - Contig-U16527-1 FC-IC01...02F (Link to Original site) FC-IC0102F 434 - - - - - - Show FC-IC0102 Library FC-IC (Link to library) Clone ...ID FC-IC0102 (Link to dictyBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U16527-1 Original site URL http://dict... (bits) Value N AB088483 |AB088483.1 Dictyostelium discoideum gene for gamete and mating-type specific prote...oducing significant alignments: (bits) Value AB088483_1( AB088483 |pid:none) Dictyostelium discoideum gmsA g

Dicty_cDB: SLC455 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available SL (Link to library) SLC455 (Link to dictyBase) - - - Contig-U16584-1 SLC455Z (Link... to Original site) - - SLC455Z 379 - - - - Show SLC455 Library SL (Link to library) Clone ID SLC455 (Link to...ycdb.biol.tsukuba.ac.jp/CSM/SL/SLC4-C/SLC455Q.Seq.d/ Representative seq. ID SLC45...5Z (Link to Original site) Representative DNA sequence >SLC455 (SLC455Q) /CSM/SL/SLC4-C/SLC455Q.Seq.d/ XXXXX...57 ) Dictyostelium discoideum slug cDNA, clone SLC469. 468 e-177 3 ( AU034549 ) Dictyostelium discoideum slug cDNA, clone SLC4

Dicty_cDB: SSI186 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available daria macrospora app gene for a... 58 3e-07 BX571864_113... 25 k... 359 e-102 AC116551_17( AC116551 |pid:none) Dictyostelium discoideum chromoso... 98 2e-19 AJ575138_1( AJ575138 |pid:none) Sor

Medicinal chemistry of adenosine, P2Y and P2X receptors.

Science.gov (United States)

Jacobson, Kenneth A; Müller, Christa E

2016-05-01

Pharmacological tool compounds are now available to define action at the adenosine (ARs), P2Y and P2X receptors. We present a selection of the most commonly used agents to study purines in the nervous system. Some of these compounds, including A1 and A3 AR agonists, P2Y1R and P2Y12R antagonists, and P2X3, P2X4 and P2X7 antagonists, are potentially of clinical use in treatment of disorders of the nervous system, such as chronic pain, neurodegeneration and brain injury. Agonists of the A2AAR and P2Y2R are already used clinically, P2Y12R antagonists are widely used antithrombotics and an antagonist of the A2AAR is approved in Japan for treating Parkinson's disease. The selectivity defined for some of the previously introduced compounds has been revised with updated pharmacological characterization, for example, various AR agonists and antagonists were deemed A1AR or A3AR selective based on human data, but species differences indicated a reduction in selectivity ratios in other species. Also, many of the P2R ligands still lack bioavailability due to charged groups or hydrolytic (either enzymatic or chemical) instability. X-ray crystallographic structures of AR and P2YRs have shifted the mode of ligand discovery to structure-based approaches rather than previous empirical approaches. The X-ray structures can be utilized either for in silico screening of chemically diverse libraries for the discovery of novel ligands or for enhancement of the properties of known ligands by chemical modification. Although X-ray structures of the zebrafish P2X4R have been reported, there is scant structural information about ligand recognition in these trimeric ion channels. In summary, there are definitive, selective agonists and antagonists for all of the ARs and some of the P2YRs; while the pharmacochemistry of P2XRs is still in nascent stages. The therapeutic potential of selectively modulating these receptors is continuing to gain interest in such fields as cancer, inflammation, pain

Dicty_cDB: VHK209 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VH (Link to library) VHK209 (Link to dictyBase) - - - Contig-U16260-1 VHK209P (Link to Original site) VHK2...09F 573 VHK209Z 765 VHK209P 1318 - - Show VHK209 Library VH (Link to library) Clone ID VHK2...e URL http://dictycdb.biol.tsukuba.ac.jp/CSM/VH/VHK2-A/VHK209Q.Seq.d/ Representative seq. ID VHK2...09P (Link to Original site) Representative DNA sequence >VHK209 (VHK209Q) /CSM/VH/VHK2-A/VHK2...Sequences producing significant alignments: (bits) Value N ( BJ446805 ) Dictyostelium discoideum cDNA clone:ddv63k2

Journal of Biosciences | Indian Academy of Sciences

Indian Academy of Sciences (India)

What history tells us XXXVII. ... Classification and expression analyses of homeobox genes from Dictyostelium discoideum .... TORC2 and eisosomes are spatially interdependent, requiring optimal level of phosphatidylinositol .... A sample of 14 schizophrenia patients and 14 healthy control subjects matched for age, sex and ...

Dicty_cDB: SSH868 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available 5 k... 329 5e-89 AC116551_17( AC116551 |pid:none) Dictyostelium discoideum chromoso... 95 2e-18 AJ575138_1( AJ575138 |pid:none) Sorda...ria macrospora app gene for a... 58 3e-07 BX571864_113(

Dicty_cDB: VHK207 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VH (Link to library) VHK207 (Link to dictyBase) - - - Contig-U16260-1 VHK207P (Link to Original site) VHK2...07F 371 VHK207Z 713 VHK207P 1064 - - Show VHK207 Library VH (Link to library) Clone ID VHK2...e URL http://dictycdb.biol.tsukuba.ac.jp/CSM/VH/VHK2-A/VHK207Q.Seq.d/ Representative seq. ID VHK2...07P (Link to Original site) Representative DNA sequence >VHK207 (VHK207Q) /CSM/VH/VHK2-A/VHK2...ducing significant alignments: (bits) Value N ( BJ446805 ) Dictyostelium discoideum cDNA clone:ddv63k21, 3'

Dicty_cDB: SLE172 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available d/ 1126 0.0 VHJ195 (VHJ195Q) /CSM/VH/VHJ1-D/VHJ195Q.Seq.d/ 1102 0.0 SLD492 (SLD492Q) /CSM/SL/SLD4-D/SLD492... 0.0 1 ( AU052538 ) Dictyostelium discoideum slug cDNA, clone SLD492. 817 0.0 1 ( C83939 ) Dictyostelium dis

Selfish DNA: a pharmaceutical perspective.

Science.gov (United States)

Winckler, T

2013-07-01

Almost 25 years ago, Theo Dingermann published the discovery of a new mobile genetic element in the unicellular microbe Dictyostelium discoideum in the journal Science. An interesting property of this new molecular parasite, the Dictyostelium Repetitive Element (DRE), was that all integrations were found approximately 50 base pairs (bp) upstream of transfer RNA (tRNA) genes in the D. discoideum genome, thus implying an active targeting mechanism to avoid the disruption of host cell genes by the retrotransposition process. Since then, the facultative multicellular "social amoeba" D. discoideum has become a popular model for analyzing complex cellular functions such as cell movement, chemotaxis, phagocytosis, and cell differentiation, important areas of biomedical research that are often hard to investigate in cells from "higher organisms" including humans. Therefore, progress in the development of methods to study Dictyostelium biology has also provoked research on transposable elements in this organism. Early work on the DRE element suggested that studying its molecular mechanism of site-specific integration might promote human gene therapy technology through the design of integrating gene transfer vectors with low intrinsic genotoxic potential. In this review article, I will briefly review the original research performed on the DRE transposable element in the Dingermann lab and report on how the emergence of genomics technologies and the development of tools to analyze de novo retrotransposition events in D. discoideum cells will expand our knowledge of DRE biology in the future.

Dicty_cDB: Contig-U01715-1 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available 0 2 ( AC116924 ) Dictyostelium discoideum chromosome 2 map 6357117... 50 0.17 1 ( FG297596 ) 1108793288766 New World... Screwworm Larvae 9387 EST... 42 0.20 2 ( FG295480 ) 1108770732782 New World...57227 Global-Ocean-Sampling_GS-27-01-01-1... 38 0.71 2 ( FG297390 ) 1108793286096 New World

Study of TiO2(1 1 0)-p(1x1), p(1x2) and p(1x3) surface structures by impact collision ion scattering spectroscopy (ICISS)

International Nuclear Information System (INIS)

Asari, E.; Souda, R.

2000-01-01

The surface structure of TiO 2 (1 1 0)-p(1x1), p(1x2) and p(1x3) were studied using impact collision ion scattering spectroscopy (ICISS). We found that (i) the height of bridging oxygen for the p(1x1) is comparative to that of bulk structure, (ii) the p(1x2) surface has the added Ti 2 O 3 unit rows proposed by Onishi et al. and also the oxygen atoms rows between Ti 2 O 3 unit rows and (iii) the p(1x3) surface is constructed with the same added Ti 2 O 3 unit rows as that in the p(1x2) surface, but the bridging oxygen rows exist between the Ti 2 O 3 unit rows

Dicty_cDB: SLB690 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available r Dp87... 45 9e-04 AC117267_7( AC117267 |pid:none) Dictyostelium discoideum chromosom... 40 0.037 AY574051_1( AY574051 |pid:none) Ict...SL (Link to library) SLB690 (Link to dictyBase) - - - Contig-U16325-1 SLB690Z (Link... to Original site) - - SLB690Z 481 - - - - Show SLB690 Library SL (Link to library) Clone ID SLB690 (Link to dict...yBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U16325-1 Original site URL http://dict...t alignments: (bits) Value N ( AF066071 ) Dictyostelium discoideum SP85 (pspB) gene, comple... 831 0.0 1 ( AC117075 ) Dict

Dicty_cDB: Contig-U05076-1 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available Dictyostelium discoideum slug cDNA, clone SSF125. 767 0.0 2 ( FG291554 ) 1108793348415 New World Screwworm E...CF-24-HW liver cDNA li... 48 0.34 1 ( FG284835 ) 1108770682807 New World Screwworm Egg 9261 ESTs C... 36 0.9

P2X1 receptors and the endothelium

Directory of Open Access Journals (Sweden)

LS Harrington

2005-03-01

Full Text Available Adenosine triphosphate (ATP is now established as a principle vaso-active mediator in the vasculature. Its actions on arteries are complex, and are mediated by the P2X and P2Y receptor families. It is generally accepted that ATP induces a bi-phasic response in arteries, inducing contraction via the P2X and P2Y receptors on the smooth muscle cells, and vasodilation via the actions of P2Y receptors located on the endothelium. However, a number of recent studies have placed P2X1 receptors on the endothelium of some arteries. The use of a specific P2X1 receptor ligand, a, b methylene ATP has demonstrated that P2X1 receptors also have a bi-functional role. The actions of ATP on P2X1 receptors is therefore dependant on its location, inducing contraction when located on the smooth muscle cells, and dilation when expressed on the endothelium, comparable to that of P2Y receptors.

P2X receptors in epithelia

DEFF Research Database (Denmark)

Leipziger, Jens Georg

2015-01-01

P2X receptors are ubiquitously expressed in all epithelial tissues but their functional roles are less well studied. Here we review the current state of knowledge by focusing on functional effects of P2X receptor in secretory and in absorptive tissues. In glandular tissue like the parotid gland...

Dicty_cDB: [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VF (Link to library) VFB585 (Link to dictyBase) - - - Contig-U09875-1 VFB585Z (Link... to Original site) - - VFB585Z 664 - - - - Show VFB585 Library VF (Link to library) Clone ID VFB585 (Link to dict...yBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U09875-1 Original site URL http://dict...Score E Sequences producing significant alignments: (bits) Value N AC116551 |AC116551.2 Dictyostelium discoi...ces producing significant alignments: (bits) Value AC116551_43( AC116551 |pid:none) Dictyostelium discoideum

Dicty_cDB: Contig-U15060-1 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available 000227 |pid:none) Bacillus cereus Q1, complete ge... 114 3e-23 B83991( B83991 ) glycolate oxidase subunit BH2730 [imported...ana interm... 56 2e-15 5 ( AF211126 ) Carsonella ruddii natural-host Bactericera cocker....psnkfvpqrlfqq*fvf tiqrkln*vllgnqvkvl*vnsqvqwlksifitfvplisrmfvslslvskvqrrl*isie lqfsissprmlplv*vlllvklgpkkdmi... la... 1074 0.0 1 ( AB000109 ) Dictyostelium discoideum mitochondrial DNA, compl... 1074 0.0 1 ( BJ412759 ) Dictyosteli...7, 3' ... 731 0.0 1 ( DQ336395 ) Dictyostelium citrinum mitochondrion, complete ge... 456 0.0 3 ( BJ387435 ) Dictyosteli

Dicty_cDB: VHK296 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VH (Link to library) VHK296 (Link to dictyBase) - - - Contig-U16260-1 VHK296P (Link to Original site) VHK2...96F 531 VHK296Z 314 VHK296P 825 - - Show VHK296 Library VH (Link to library) Clone ID VHK2... URL http://dictycdb.biol.tsukuba.ac.jp/CSM/VH/VHK2-D/VHK296Q.Seq.d/ Representative seq. ID VHK2...96P (Link to Original site) Representative DNA sequence >VHK296 (VHK296Q) /CSM/VH/VHK2-D/VHK2... 5' ... 993 0.0 1 ( BJ427875 ) Dictyostelium discoideum cDNA clone:ddv63k24, 5' ... 993 0.0 1 ( BJ427874 ) D

Dicty_cDB: VFK242 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VF (Link to library) VFK242 (Link to dictyBase) - - - Contig-U16260-1 VFK242P (Link to Original site) VFK2...42F 606 VFK242Z 292 VFK242P 878 - - Show VFK242 Library VF (Link to library) Clone ID VFK2... URL http://dictycdb.biol.tsukuba.ac.jp/CSM/VF/VFK2-B/VFK242Q.Seq.d/ Representative seq. ID VFK2...42P (Link to Original site) Representative DNA sequence >VFK242 (VFK242Q) /CSM/VF/VFK2-B/VFK2...eum cDNA clone:ddv63n23, 5' ... 1142 0.0 1 ( BJ427875 ) Dictyostelium discoideum cDNA clone:ddv63k24, 5' ...

Dicty_cDB: SLH243 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available SL (Link to library) SLH243 (Link to dictyBase) - - - Contig-U15469-1 SLH243P (Link to Original site) SLH2...43F 436 SLH243Z 369 SLH243P 805 - - Show SLH243 Library SL (Link to library) Clone ID SLH2... URL http://dictycdb.biol.tsukuba.ac.jp/CSM/SL/SLH2-B/SLH243Q.Seq.d/ Representative seq. ID SLH2...43P (Link to Original site) Representative DNA sequence >SLH243 (SLH243Q) /CSM/SL/SLH2-B/SLH2...SA605. 785 0.0 1 ( AU062023 ) Dictyostelium discoideum slug cDNA, clone SLH243. 785 0.0 1 ( BJ416791 ) Dicty

P2X7 receptors in satellite glial cells mediate high functional expression of P2X3 receptors in immature dorsal root ganglion neurons

Directory of Open Access Journals (Sweden)

Chen Yong

2012-02-01

Full Text Available Abstract Background The purinergic P2X3 receptor (P2X3R expressed in the dorsal root ganglion (DRG sensory neuron and the P2X7 receptor (P2X7R expressed in the surrounding satellite glial cell (SGC are two major receptors participating in neuron-SGC communication in adult DRGs. Activation of P2X7Rs was found to tonically reduce the expression of P2X3Rs in DRGs, thus inhibiting the abnormal pain behaviors in adult rats. P2X receptors are also actively involved in sensory signaling in developing rodents. However, very little is known about the developmental change of P2X7Rs in DRGs and the interaction between P2X7Rs and P2X3Rs in those animals. We therefore examined the expression of P2X3Rs and P2X7Rs in postnatal rats and determined if P2X7R-P2X3R control exists in developing rats. Findings We immunostained DRGs of immature rats and found that P2X3Rs were expressed only in neurons and P2X7Rs were expressed only in SGCs. Western blot analyses indicated that P2X3R expression decreased while P2X7R expression increased with the age of rats. Electrophysiological studies showed that the number of DRG neurons responding to the stimulation of the P2XR agonist, α,β-meATP, was higher and the amplitudes of α,β-meATP-induced depolarizations were larger in immature DRG neurons. As a result, P2X3R-mediated flinching responses were much more pronounced in immature rats than those found in adult rats. When we reduced P2X7R expression with P2X7R-siRNA in postnatal and adult rats, P2X3R-mediated flinch responses were greatly enhanced in both rat populations. Conclusions These results show that the P2X7R expression increases as rats age. In addition, P2X7Rs in SGCs exert inhibitory control on the P2X3R expression and function in sensory neurons of immature rats, just as observed in adult rats. Regulation of P2X7R expression is likely an effective way to control P2X3R activity and manage pain relief in infants.

Dicty_cDB: [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VF (Link to library) VFB611 (Link to dictyBase) - - - Contig-U16239-1 VFB611P (Link... to Original site) VFB611F 650 VFB611Z 704 VFB611P 1354 - - Show VFB611 Library VF (Link to library) Clone ID VFB611 (Link to dict...yBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U16239-1 Original site URL http://dict...ant alignments: (bits) Value N AC116989 |AC116989.2 Dictyostelium discoideum chro...mosome 2 map complement(3527391-3470188) strain AX4, complete sequence. 42 5e-10 9 AC116987 |AC116987.2 Dict

Dicty_cDB: [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VF (Link to library) VFB574 (Link to dictyBase) - - - Contig-U16382-1 VFB574P (Link... to Original site) VFB574F 508 VFB574Z 686 VFB574P 1194 - - Show VFB574 Library VF (Link to library) Clone ID VFB574 (Link to dict...yBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U16382-1 Original site URL http://dict...uences producing significant alignments: (bits) Value N AC116957 |AC116957.2 Dictyostelium discoideum chromo...some 2 map 1685067-2090751 strain AX4, complete sequence. 1352 0.0 4 AC116986 |AC116986.2 Dict

Immunocytochemical analysis of P2X2 in rat circumvallate taste buds.

Science.gov (United States)

Yang, Ruibiao; Montoya, Alana; Bond, Amanda; Walton, Jenna; Kinnamon, John C

2012-05-23

Our laboratory has shown that classical synapses and synaptic proteins are associated with Type III cells. Yet it is generally accepted that Type II cells transduce bitter, sweet and umami stimuli. No classical synapses, however, have been found associated with Type II cells. Recent studies indicate that the ionotropic purinergic receptors P2X2/P2X3 are present in rodent taste buds. Taste nerve processes express the ionotropic purinergic receptors (P2X2/P2X3). P2X2/P2X3(Dbl-/-) mice are not responsive to sweet, umami and bitter stimuli, and it has been proposed that ATP acts as a neurotransmitter in taste buds. The goal of the present study is to learn more about the nature of purinergic contacts in rat circumvallate taste buds by examining immunoreactivity to antisera directed against the purinergic receptor P2X2. P2X2-like immunoreactivity is present in intragemmal nerve processes in rat circumvallate taste buds. Intense immunoreactivity can also be seen in the subgemmal nerve plexuses located below the basal lamina. The P2X2 immunoreactive nerve processes also display syntaxin-1-LIR. The immunoreactive nerves are in close contact with the IP(3)R3-LIR Type II cells and syntaxin-1-LIR and/or 5-HT-LIR Type III cells. Taste cell synapses are observed only from Type III taste cells onto P2X2-LIR nerve processes. Unusually large, "atypical" mitochondria in the Type II taste cells are found only at close appositions with P2X2-LIR nerve processes. P2X2 immunogold particles are concentrated at the membranes of nerve processes at close appositions with taste cells. Based on our immunofluorescence and immunoelectron microscopical studies we believe that both perigemmal and most all intragemmal nerve processes display P2X2-LIR. Moreover, colloidal gold immunoelectron microscopy indicates that P2X2-LIR in nerve processes is concentrated at sites of close apposition with Type II cells. This supports the hypothesis that ATP may be a key neurotransmitter in taste transduction

Dicty_cDB: VFB113 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VF (Link to library) VFB113 (Link to dictyBase) - - - Contig-U16478-1 VFB113P (Link... to Original site) VFB113F 584 VFB113Z 643 VFB113P 1227 - - Show VFB113 Library VF (Link to library) Clone ID VFB113 (Link to dict...yBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U16478-1 Original site URL http://dict...rlstttrlptttrlptttrlstttr lptttrlptttrlptttrlptttrlsttrlstttrlstswctswctswictrygswissr llcwynhs*l*t*c*sfkksn...sequence. 42 5e-29 8 U03413 |U03413.1 Dictyostelium discoideum AX2 calcium binding protein mRNA, complete cd

Dicty_cDB: [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VF (Link to library) VFC231 (Link to dictyBase) - - - Contig-U16593-1 VFC231P (Link... to Original site) VFC231F 428 VFC231Z 202 VFC231P 630 - - Show VFC231 Library VF (Link to library) Clone ID VFC231 (Link to dict...yBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U16593-1 Original site URL http://dict...amoeba histolytica genomic, DNA sequence. 48 0.13 1 AC116984 |AC116984.2 Dictyostelium discoideum chromosome...mbrane 4.0 %: vesicles of secretory system 4.0 %: extracellular, including cell wall >> predict

The Amoebal MAP Kinase Response to Legionella pneumophila Is Regulated by DupA

OpenAIRE

Li, Zhiru; Dugan, Aisling S.; Bloomfield, Gareth; Skelton, Jason; Ivens, Alasdair; Losick, Vicki; Isberg, Ralph R.

2009-01-01

The amoeba Dictyostelium discoideum can support replication of Legionella pneumophila. Here we identify the dupA gene, encoding a putative tyrosine kinase/dual-specificity phosphatase, in a screen for D. discoideum mutants altered in allowing L. pneumophila intracellular replication. Inactivation of dupA resulted in depressed L. pneumophila growth and sustained hyperphosphorylation of the amoebal MAP kinase ERK1, consistent with loss of a phosphatase activity. Bacterial challenge of wild-type...

Chronic administration of the selective P2X3, P2X2/3 receptor antagonist, A-317491, transiently attenuates cancer-induced bone pain in mice

DEFF Research Database (Denmark)

Hansen, Rikke Rie; Nasser, Arafat; Falk, Sarah

2012-01-01

The purinergic P2X3 and P2X2/3 receptors are in the peripheral nervous system almost exclusively confined to afferent sensory neurons, where they are found both at peripheral and central synapses. The P2X3 receptor is implicated in both neuropathic and inflammatory pain. However, the role of the ......X3 receptor in chronic cancer-induced bone pain is less known. Here we investigated the effect of systemic acute and chronic administration of the selective P2X3, P2X2/3 receptor antagonist (5-[[[(3-Phenoxyphenyl)methyl][(1S)-1,2,3,4-tetrahydro-1-naphthalenyl]amino]carbonyl]-1...

Immunocytochemical analysis of P2X2 in rat circumvallate taste buds

Directory of Open Access Journals (Sweden)

Yang Ruibiao

2012-05-01

Full Text Available Abstract Background Our laboratory has shown that classical synapses and synaptic proteins are associated with Type III cells. Yet it is generally accepted that Type II cells transduce bitter, sweet and umami stimuli. No classical synapses, however, have been found associated with Type II cells. Recent studies indicate that the ionotropic purinergic receptors P2X2/P2X3 are present in rodent taste buds. Taste nerve processes express the ionotropic purinergic receptors (P2X2/P2X3. P2X2/P2X3Dbl−/− mice are not responsive to sweet, umami and bitter stimuli, and it has been proposed that ATP acts as a neurotransmitter in taste buds. The goal of the present study is to learn more about the nature of purinergic contacts in rat circumvallate taste buds by examining immunoreactivity to antisera directed against the purinergic receptor P2X2. Results P2X2-like immunoreactivity is present in intragemmal nerve processes in rat circumvallate taste buds. Intense immunoreactivity can also be seen in the subgemmal nerve plexuses located below the basal lamina. The P2X2 immunoreactive nerve processes also display syntaxin-1-LIR. The immunoreactive nerves are in close contact with the IP3R3-LIR Type II cells and syntaxin-1-LIR and/or 5-HT-LIR Type III cells. Taste cell synapses are observed only from Type III taste cells onto P2X2-LIR nerve processes. Unusually large, “atypical” mitochondria in the Type II taste cells are found only at close appositions with P2X2-LIR nerve processes. P2X2 immunogold particles are concentrated at the membranes of nerve processes at close appositions with taste cells. Conclusions Based on our immunofluorescence and immunoelectron microscopical studies we believe that both perigemmal and most all intragemmal nerve processes display P2X2-LIR. Moreover, colloidal gold immunoelectron microscopy indicates that P2X2-LIR in nerve processes is concentrated at sites of close apposition with Type II cells. This supports the hypothesis

PsB multiprotein complex of Dictyostelium discoideum. Demonstration of cellulose binding activity and order of protein subunit assembly.

Science.gov (United States)

McGuire, V; Alexander, S

1996-06-14

The differentiated spores of Dictyostelium are surrounded by an extracellular matrix, the spore coat, which protects them from environmental factors allowing them to remain viable for extended periods of time. This presumably is a major evolutionary advantage. This unique extracellular matrix is composed of cellulose and glycoproteins. Previous work has shown that some of these spore coat glycoproteins exist as a preassembled multiprotein complex (the PsB multiprotein complex) which is stored in the prespore vesicles (Watson, N., McGuire, V., and Alexander, S (1994) J. Cell Sci. 107, 2567-2579). Later in development, the complex is synchronously secreted from the prespore vesicles and incorporated into the spore coat. We now have shown that the PsB complex has a specific in vitro cellulose binding activity. The analysis of mutants lacking individual subunits of the PsB complex revealed the relative order of assembly of the subunit proteins and demonstrated that the protein subunits must be assembled for cellulose binding activity. These results provide a biochemical explanation for the localization of this multiprotein complex in the spore coat.

Journal of Biosciences | Indian Academy of Sciences

Indian Academy of Sciences (India)

A model for cell type localization in the migrating slug of Dictyostelium discoideum based on differential ... a progressive maturation of chemotactic properties during the transdifferentiation of slug cell types. ... Journal of Biosciences | News ...

Dicty_cDB: Contig-U15582-1 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available ) EST1120673 Aquilegia cDNA library Aquilegia formo... 36 0.36 3 ( AC115612 ) Dictyostelium discoideum chrom... cDNA clone ... 46 4.1 1 ( BX858993 ) AGENAE Rainbow trout normalized testis library (t... 46 4.1 1 ( BF0889...discoideum chromosome 2 map 2567470... 40 0.11 12 ( AL844509 ) Plasmodium falciparum chromosome 13. 38 0.15 17 ( EU016597 ) Unculture...EST1196545 Aquilegia cDNA library Aquilegia formo... 36 0.28 3 ( DT766291 ) EST1200140 Aquilegia cDNA libr...ary Aquilegia formo... 36 0.29 3 ( DT729293 ) EST1163143 Aquilegia cDNA library Aqu

P2X4: A fast and sensitive purinergic receptor

Directory of Open Access Journals (Sweden)

Jaanus Suurväli

2017-10-01

Full Text Available Extracellular nucleotides have been recognized as important mediators of activation, triggering multiple responses via plasma membrane receptors known as P2 receptors. P2 receptors comprise P2X ionotropic receptors and G protein-coupled P2Y receptors. P2X receptors are expressed in many tissues, where they are involved in a number of functions including synaptic transmission, muscle contraction, platelet aggregation, inflammation, macrophage activation, differentiation and proliferation, neuropathic and inflammatory pain. P2X4 is one of the most sensitive purinergic receptors (at nanomolar ATP concentrations, about one thousand times more than the archetypal P2X7. P2X4 is widely expressed in central and peripheral neurons, in microglia, and also found in various epithelial tissues and endothelial cells. It localizes on the plasma membrane, but also in intracellular compartments. P2X4 is preferentially localized in lysosomes, where it is protected from proteolysis by its glycosylation. High ATP concentration in the lysosomes does not activate P2X4 at low pH; P2X4 gets activated by intra-lysosomal ATP only in its fully dissociated tetra-anionic form, when the pH increases to 7.4. Thus, P2X4 is functioning as a Ca2+-channel after the fusion of late endosomes and lysosomes. P2X4 modulates major neurotransmitter systems and regulates alcohol-induced responses in microglia. P2X4 is one of the key receptors mediating neuropathic pain. However, injury-induced upregulation of P2X4 expression is gender dependent and plays a key role in pain difference between males and females. P2X4 is also involved in inflammation. Extracellular ATP being a pro-inflammatory molecule, P2X4 can trigger inflammation in response to high ATP release. It is therefore involved in multiple pathologies, like post-ischemic inflammation, rheumatoid arthritis, airways inflammation in asthma, neurodegenerative diseases and even metabolic syndrome. Although P2X4 remains poorly

Dicty_cDB: SLH294 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available SL (Link to library) SLH294 (Link to dictyBase) - - - Contig-U16466-1 SLH294E (Link... to Original site) - - - - - - SLH294E 393 Show SLH294 Library SL (Link to library) Clone ID SLH294 (Link to...ycdb.biol.tsukuba.ac.jp/CSM/SL/SLH2-D/SLH294Q.Seq.d/ Representative seq. ID SLH29...4E (Link to Original site) Representative DNA sequence >SLH294 (SLH294Q) /CSM/SL/SLH2-D/SLH294Q.Seq.d/ CGATA...stelium discoideum slug cDNA, clone SLH294. 666 0.0 1 ( AU034499 ) Dictyostelium discoideum slug cDNA, clone

Desynchronization of cells on the developmental path triggers the formation of spiral waves of cAMP during Dictyostelium aggregation

OpenAIRE

Lauzeral, Jacques; Halloy, José; Goldbeter, Albert

1997-01-01

Whereas it is relatively easy to account for the formation of concentric (target) waves of cAMP in the course of Dictyostelium discoideum aggregation after starvation, the origin of spiral waves remains obscure. We investigate a physiologically plausible mechanism for the spontaneous formation of spiral waves of cAMP in D. discoideum. The scenario relies on the developmental path associated with the continuous changes in the activity of enzymes such as adenylate cyclase and phosphodiesterase ...

Dicty_cDB: SLC466 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available SL (Link to library) SLC466 (Link to dictyBase) - - - Contig-U16381-1 SLC466Z (Link... to Original site) - - SLC466Z 427 - - - - Show SLC466 Library SL (Link to library) Clone ID SLC466 (Link to...ycdb.biol.tsukuba.ac.jp/CSM/SL/SLC4-C/SLC466Q.Seq.d/ Representative seq. ID SLC46...6Z (Link to Original site) Representative DNA sequence >SLC466 (SLC466Q) /CSM/SL/SLC4-C/SLC466Q.Seq.d/ XXXXX... AU034556 ) Dictyostelium discoideum slug cDNA, clone SLC466. 176 2e-77 2 ( AU033496 ) Dictyostelium discoid

Dicty_cDB: SLA508 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available e-101 SLD569 (SLD569Q) /CSM/SL/SLD5-C/SLD569Q.Seq.d/ 250 1e-65 SLD492 (SLD492Q) /CSM/SL/SLD4-D/SLD492...oideum slug cDNA, clone SLD569. 250 2e-62 1 ( AU052538 ) Dictyostelium discoideum slug cDNA, clone SLD492. 7

P2X receptors, sensory neurons and pain.

Science.gov (United States)

Bele, Tanja; Fabbretti, Elsa

2015-01-01

Pain represents a very large social and clinical problem since the current treatment provides insufficient pain relief. Plasticity of pain receptors together with sensitisation of sensory neurons, and the role of soluble mediators released from non-neuronal cells render difficult to understand the spatial and temporal scale of pain development, neuronal responses and disease progression. In pathological conditions, ATP is one of the most powerful mediators that activates P2X receptors that behave as sensitive ATP-detectors, such as neuronal P2X3 receptor subtypes and P2X4 and P2X7 receptors expressed on non-neuronal cells. Dissecting the molecular mechanisms occurring in sensory neurons and in accessory cells allows to design appropriate tissue- and cell- targeted approaches to treat chronic pain.

Interaction of GABAA receptors with purinergic P2X2 receptors

International Nuclear Information System (INIS)

Shrivastava, A.

2010-01-01

GABA A Rs in the spinal cord are evolving as an important target for drug development against pain. Purinergic P2X 2 Rs are also expressed in spinal cord neurons and are known to cross-talk with GABA A Rs. Here we investigated a possible 'dynamic' interaction between GABA A Rs and P2X 2 Rs using co-immunoprecipitation and FRET studies in HEK cells along with co-localization and single particle tracking studies in spinal cord neurons. Our results suggest that a significant proportion of P2X 2 Rs forms a transient complex with GABA A Rs inside the cell, thus stabilizing these receptors and using them for co-trafficking to the cell surface. P2X 2 Rs and GABA A Rs are then co-inserted into the cell membrane and are primarily located extra-synaptically. Furthermore, agonist induced activation of P2X 2 Rs results in disassembly of the receptor complex and destabilization of GABA A Rs whereas P2X 2 Rs are stabilized and form larger clusters. Antagonist-induced blocking of P2XRs results in co-stabilization of this receptor complex at the cell surface. These results suggest a novel mechanism where association of P2XRs with other receptors could be used for specific targeting to the neuronal membrane, thus providing an extrasynaptic receptor reserve that could regulate the excitability of neurons. We further conclude that blocking the excitatory activity of excessively released ATP under diseased state by P2XR antagonists could simultaneously enhance synaptic inhibition mediated by GABA A Rs.(author) (author) [de

A model for cell type localization in the migrating slug of ...

Indian Academy of Sciences (India)

PRAKASH

. Localization of the three major cell types within the migrating slug stage is a dynamic process (Sternfeld 1992;. A model for cell type localization in the migrating slug of Dictyostelium discoideum based on differential chemotactic sensitivity to ...

Structural and Molecular Modeling Features of P2X Receptors

Directory of Open Access Journals (Sweden)

Luiz Anastacio Alves

2014-03-01

Full Text Available Currently, adenosine 5'-triphosphate (ATP is recognized as the extracellular messenger that acts through P2 receptors. P2 receptors are divided into two subtypes: P2Y metabotropic receptors and P2X ionotropic receptors, both of which are found in virtually all mammalian cell types studied. Due to the difficulty in studying membrane protein structures by X-ray crystallography or NMR techniques, there is little information about these structures available in the literature. Two structures of the P2X4 receptor in truncated form have been solved by crystallography. Molecular modeling has proven to be an excellent tool for studying ionotropic receptors. Recently, modeling studies carried out on P2X receptors have advanced our knowledge of the P2X receptor structure-function relationships. This review presents a brief history of ion channel structural studies and shows how modeling approaches can be used to address relevant questions about P2X receptors.

Dicty_cDB: SFF154 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available SF (Link to library) SFF154 (Link to dictyBase) - - - Contig-U15074-1 SFF154P (Link... to Original site) SFF154F 132 SFF154Z 514 SFF154P 646 - - Show SFF154 Library SF (Link to library) Clone ID SFF154 (Link to dict...yBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U15074-1 Original site URL http://dict...22, genomic survey sequence. 48 3e-09 3 AC116921 |AC116921.2 Dictyostelium discoi...deum chromosome 2 map 4624505-4657775 strain AX4, complete sequence. 44 4e-09 7 BQ096682 |BQ096682.1 IfHdk00151 Ict

Dicty_cDB: [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VF (Link to library) VFC196 (Link to dictyBase) - - - Contig-U13856-1 VFC196P (Link... to Original site) VFC196F 487 VFC196Z 556 VFC196P 1043 - - Show VFC196 Library VF (Link to library) Clone ID VFC196 (Link to dict...yBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U13856-1 Original site URL http://dict...te 2004.12.25 Homology vs DNA Score E Sequences producing significant alignments: (bits) Value N U72746 |U72746.1 Dict...13. 36 0.053 5 AC116984 |AC116984.2 Dictyostelium discoideum chromosome 2 map 2567470-3108875 strain AX4, co

Dicty_cDB: VHA340 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available _1( AY698035 |pid:none) Desmognathus marmoratus isolate 69... 33 5.9 AC116957_68( AC116957 |pid:none) Dictyo...stelium discoideum chromoso... 33 7.6 AY612344_1( AY612344 |pid:none) Desmognathus marmoratus voucher KH...

Dicty_cDB: VHI760 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available 7( AC116551 |pid:none) Dictyostelium discoideum chromoso... 79 3e-13 AJ575138_1( AJ575138 |pid:none) Sorda...ria macrospora app gene for a... 50 1e-04 BX571864_113( BX571864 |pid:none) Photorh

Dicty_cDB: [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VF (Link to library) VFB789 (Link to dictyBase) - - - Contig-U16455-1 VFB789P (Link... to Original site) VFB789F 468 VFB789Z 735 VFB789P 1203 - - Show VFB789 Library VF (Link to library) Clone ID VFB789 (Link to dict...yBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U16455-1 Original site URL http://dict...oideum EF1-II gene for elongation factor 1 alpha. 1338 0.0 2 AF016242 |AF016242.1 Dictyostelium discoideum p...m_ : 1.00 36.0 %: cytoplasmic 36.0 %: nuclear 12.0 %: cytoskeletal 8.0 %: vacuolar 4.0 %: mitochondrial 4.0 %: peroxisomal >> predict

Dicty_cDB: SSE149 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available SS (Link to library) SSE149 (Link to dictyBase) - - - Contig-U01658-1 | Contig-U165... library) Clone ID SSE149 (Link to dictyBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U016...58-1 | Contig-U16509-1 Original site URL http://dictycdb.biol.tsukuba.ac.jp/CSM/S...g significant alignments: (bits) Value N D16417 |D16417.1 Dictyostelium discoideum mRNA. 64 1e-17 2 AC100496...egans clone T13G4, complete sequence. 36 5.0 2 BM028890 |BM028890.1 IpSkn01670 Skin cDNA library Ictalurus p

Dicty_cDB: Contig-U04940-1 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available iscoideum chromosome 2 map 3622643... 32 0.081 12 ( GE806545 ) EST_scau_evk_971547 scauevk mixed_tissue Sebaste...s... 40 0.092 2 ( EW978238 ) EST_sras_evg_777463 srasevg mixed_tissue Sebastes... 40 0.093 2 ( EW984986 )... EST_sras_evg_777879 srasevg mixed_tissue Sebastes... 40 0.095 2 ( EW985351 ) EST..._sras_evg_778065 srasevg mixed_tissue Sebastes... 40 0.096 2 ( AC116305 ) Dictyostelium discoideum chromosom

Post-translational regulation of P2X receptor channels: modulation by phospholipids

Directory of Open Access Journals (Sweden)

Louis-Philippe eBernier

2013-11-01

Full Text Available P2X receptor channels mediate fast excitatory signaling by ATP and play major roles in sensory transduction, neuro-immune communication and inflammatory response. P2X receptors constitute a gene family of calcium-permeable ATP-gated cation channels therefore the regulation of P2X signaling is critical for both membrane potential and intracellular calcium homeostasis. Phosphoinositides (PIPn are anionic signaling phospholipids that act as functional regulators of many types of ion channels. Direct PIPn binding was demonstrated for several ligand- or voltage-gated ion channels, however no generic motif emerged to accurately predict lipid-protein binding sites. This review presents what is currently known about the modulation of the different P2X subtypes by phospholipids and about critical determinants underlying their sensitivity to PIPn levels in the plasma membrane.All functional mammalian P2X subtypes tested, with the notable exception of P2X5, have been shown to be positively modulated by PIPn, i.e. homomeric P2X1, P2X2, P2X3, P2X4, and P2X7, as well as heteromeric P2X1/5 and P2X2/3 receptors. Based on various results reported on the aforementioned subtypes including mutagenesis of the prototypical PIPn-sensitive P2X4 and PIPn-insensitive P2X5 receptor subtypes, an increasing amount of functional, biochemical and structural evidence converges on the modulatory role of a short polybasic domain located in the proximal C-terminus of P2X subunits. This linear motif, semi-conserved in the P2X family, seems necessary and sufficient for encoding direct modulation of ATP-gated channels by PIPn. Furthermore, the physiological impact of the regulation of ionotropic purinergic responses by phospholipids on pain pathways was recently revealed in the context of native crosstalks between phospholipase C-linked metabotropic receptors and P2X receptor channels in DRG sensory neurons and microglia.

Chemotaxis in the cellular slime molds : I. The effect of temperature

NARCIS (Netherlands)

Konijn, Theo M.

1965-01-01

The effect of temperature on chemotaxis in the cellular slime mold Dictyostelium discoideum has been studied by incubating small populations of washed myxamoebae at different temperatures. Droplets containing a cell suspension of known density were deposited on a hydrophobic agar surface. The

Dicty_cDB: CFG115 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available CF (Link to library) CFG115 (Link to dictyBase) - - - Contig-U03237-1 CFG115E (Link...) Clone ID CFG115 (Link to dictyBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U03237-1 Ori...k*NNMNGFLKSQLENKTNATTTTTTRYCNNDESCNDENLCTN EMCDPVIGCIYENISCDDDNGCTKDFCDPLTGCFHKRVVCDDNDKCTDNICNPETGTCSF RRRICT...TTTRYCNNDESCNDENLCTN EMCDPVIGCIYENISCDDDNGCTKDFCDPLTGCFHKRVVCDDNDKCTDNICNPETGTCSF RRRICTDNNDCTMDWCNQLTGECVYQ...ng significant alignments: (bits) Value N AC116984 |AC116984.2 Dictyostelium discoideum chromosome 2 map 256

Analytic derivation of the leading-order gluon distribution function G(x,Q2)=xg(x,Q2) from the proton structure function F2p(x,Q2)

International Nuclear Information System (INIS)

Block, Martin M.; Durand, Loyal; McKay, Douglas W.

2008-01-01

We derive a second-order linear differential equation for the leading-order gluon distribution function G(x,Q 2 )=xg(x,Q 2 ) which determines G(x,Q 2 ) directly from the proton structure function F 2 p (x,Q 2 ). This equation is derived from the leading-order evolution equation for F 2 p (x,Q 2 ), and does not require knowledge of either the individual quark distributions or the gluon evolution equation. Given an analytic expression that successfully reproduces the known experimental data for F 2 p (x,Q 2 ) in a domain x min (Q 2 )≤x≤x max (Q 2 ), Q min 2 ≤Q 2 ≤Q max 2 of the Bjorken variable x and the virtuality Q 2 in deep inelastic scattering, G(x,Q 2 ) is uniquely determined in the same domain. We give the general solution and illustrate the method using the recently proposed Froissart-bound-type parametrization of F 2 p (x,Q 2 ) of E. L. Berger, M. M. Block and C.-I. Tan [Phys. Rev. Lett. 98, 242001 (2007)]. Existing leading-order gluon distributions based on power-law descriptions of individual parton distributions agree roughly with the new distributions for x > or approx. 10 -3 as they should, but are much larger for x -3 .

Dicty_cDB: Contig-U04013-1 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available pid:none) Flavobacterium psychrophilum JIP... 98 3e-19 EU678894_1( EU678894 |pid:none) Bacillus intermedius ...46 3e-04 2 ( EH672780 ) CCIL10412.b1_G11.ab1 CCI(LMS) chicory Cichorium i... 46 3e-04 2 ( EJ368820 ) 1092963715450 Global-Ocean-Sampl...41 |pid:none) Sulfolobus solfataricus P2, com... 89 2e-16 AH1770( AH1770 ) conserved hypothetical protein lin2710 [imported...AY653733 ) Acanthamoeba polyphaga mimivirus, complete genome. 62 3e-05 1 ( EH678404 ) CCIL3410.b1_D14.ab1 CC...ngs S... 44 6.8 1 >( AC116963 ) Dictyostelium discoideum chromosome 2 map 4657875-4914984 strain AX4, compl

Dicty_cDB: VFH641 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VF (Link to library) VFH641 (Link to dictyBase) - - - Contig-U16151-1 VFH641E (Link... Clone ID VFH641 (Link to dictyBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U16151-1 Original site URL http://dict...KLLGVQTQAKQTRATWKIVVGDGAGAVTFKLATNGGTTEGDFTTTLTSKVLS GSDPKEVGTYYMDVTVPTGTTCTGTNGICT...KLLGVQTQAKQTRATWKIVVGDGAGAVTFKLATNGGTTEGDFTTTLTSKVLS GSDPKEVGTYYMDVTVPTGTTCTGTNGICT...nificant alignments: (bits) Value N AC116984 |AC116984.2 Dictyostelium discoideum

TRE5-A retrotransposition profiling reveals putative RNA polymerase III transcription complex binding sites on the Dictyostelium extrachromosomal rDNA element.

Directory of Open Access Journals (Sweden)

Thomas Spaller

Full Text Available The amoeba Dictyostelium discoideum has a haploid genome in which two thirds of the DNA encodes proteins. Consequently, the space available for selfish mobile elements to expand without excess damage to the host genome is limited. The non-long terminal repeat retrotransposon TRE5-A maintains an active population in the D. discoideum genome and apparently adapted to this gene-dense environment by targeting positions ~47 bp upstream of tRNA genes that are devoid of protein-coding regions. Because only ~24% of tRNA genes are associated with a TRE5-A element in the reference genome, we evaluated whether TRE5-A retrotransposition is limited to this subset of tRNA genes. We determined that a tagged TRE5-A element (TRE5-Absr integrated at 384 of 405 tRNA genes, suggesting that expansion of the current natural TRE5-A population is not limited by the availability of targets. We further observed that TRE5-Absr targets the ribosomal 5S gene on the multicopy extrachromosomal DNA element that carries the ribosomal RNA genes, indicating that TRE5-A integration may extend to the entire RNA polymerase III (Pol III transcriptome. We determined that both natural TRE5-A and cloned TRE5-Absr retrotranspose to locations on the extrachromosomal rDNA element that contain tRNA gene-typical A/B box promoter motifs without displaying any other tRNA gene context. Based on previous data suggesting that TRE5-A targets tRNA genes by locating Pol III transcription complexes, we propose that A/B box loci reflect Pol III transcription complex assembly sites that possess a function in the biology of the extrachromosomal rDNA element.

Fulltext PDF

Indian Academy of Sciences (India)

SEARCHU

Molecular mechanisms underlying thermal adaptation of xeric animals. 489. Specific and unspecific responses of plants to cold and drought stress. 501. Adaptive ... Arachidonic Acid is a chemoattractant for Dictyostelium discoideum cells. 1281 .... Contribution of root to soil respiration and carbon balance in disturbed and ...

Dicty_cDB: [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VF (Link to library) VFB534 (Link to dictyBase) - - - Contig-U15835-1 VFB534P (Link... to Original site) VFB534F 523 VFB534Z 467 VFB534P 990 - - Show VFB534 Library VF (Link to library) Clone ID VFB534 (Link to dict...yBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U15835-1 Original site URL http://dict... Homology vs DNA Score E Sequences producing significant alignments: (bits) Value N AF025951 |AF025951.1 Dict...yostelium discoideum heat-shock cognate protein 70 (hsc70) mRNA, complete cds. 1037 0.0 3 AC115684 |AC115684.2 Dict

The role of P2X receptors in bone biology.

Science.gov (United States)

Jørgensen, N R; Syberg, S; Ellegaard, M

2015-01-01

Bone is a highly dynamic organ, being constantly modeled and remodeled in order to adapt to the changing need throughout life. Bone turnover involves the coordinated actions of bone formation and bone degradation. Over the past decade great effort has been put into the examination of how P2X receptors regulate bone metabolism and especially for the P2X7 receptor an impressive amount of evidence has now documented its expression in osteoblasts, osteoclasts, and osteocytes as well as important functional roles in proliferation, differentiation, and function of the cells of bone. Key evidence has come from studies on murine knockout models and from pharmacologic studies on cells and animals. More recently, the role of P2X receptors in human bone diseases has been documented. Loss-of-functions polymorphisms in the P2X7 receptorare associated with bone loss and increased fracture risk. Very recently a report from a genetic study in multiple myeloma demonstrated that decreased P2X7 receptor function was associated with increased risk of developing multiple myeloma. In contrast, the risk of developing myeloma bone disease and subsequent vertebral fractures was increased in subjects carrying P2X7 receptor gain-of-function alleles as compared to subjects only carrying loss-of-function or normal functioning alleles. It is evident that P2X receptors are important in regulating bone turnover and maintaining bone mass, and thereby holding great potential as novel drug targets for treatment of bone diseases. However, further research is needed before we fully understand the roles and effects of P2X receptors in bone.

Localization of P2X receptor subtypes 2, 3 and 7 in human urinary bladder.

Science.gov (United States)

Svennersten, Karl; Hallén-Grufman, Katarina; de Verdier, Petra J; Wiklund, N Peter; Poljakovic, Mirjana

2015-08-08

Voiding dysfunctions are a common problem that has a severe negative impact on the quality of life. Today there is a need for new drug targets for these conditions. The role of ATP receptors in bladder physiology has been studied for some time, primarily in animal models. The aim of this work is to investigate the localization of the ATP receptors P2X2, P2X3 and P2X7 and their colocalization with vimentin and actin in the human urinary bladder. Immunohistochemical analysis was conducted on full-thickness bladder tissues from fundus and trigonum collected from 15 patients undergoing open radical cystectomy due to chronic cystitis, bladder cancer or locally advanced prostate cancer. Colocalization analyses were performed between the three different P2X subtypes and the structural proteins vimentin and actin. Specimens were examined using epifluorescence microscopy and correlation coefficients were calculated for each costaining as well as the mean distance from the laminin positive basal side of the urothelium to the vimentin positive cells located in the suburothelium. P2X2 was expressed in vimentin positive cells located in the suburothelium. Less distinct labelling of P2X2 was also observed in actin positive smooth muscle cells and in the urothelium. P2X3 was expressed in vimentin positive cells surrounding the smooth muscle, and in vimentin positive cells located in the suburothelium. Weaker P2X3 labelling was seen in the urothelium. P2X7 was expressed in the smooth muscle cells and the urothelium. In the suburothelium, cells double positive for P2X2 and vimentin where located closer to the urothelium while cells double positive for P2X3 and vimentin where located further from the urothelium. The results from this study demonstrate that there is a significant difference in the expression of the purinergic P2X2, P2X3 and P2X7 receptors in the different histological layers of the human urinary bladder.

Differential Role of Poly(ADP-ribose polymerase in D. discoideum growth and development

Directory of Open Access Journals (Sweden)

Begum Rasheedunnisa

2011-03-01

Full Text Available Abstract Background Poly(ADP-ribose polymerase is evolutionarily conserved as a responder to various forms of stress. Though PARP's role in cell death is well addressed, its role in development and multicellularity is still an enigma. We have previously reported the role of PARP in oxidative stress induced delayed development of D. discoideum. Results In the current study we highlight the involvement of PARP during D. discoideum development. Oxidative stress affects expression of aca and cAR1 thus affecting aggregation. Although parp expression is not affected during oxidative stress but it is involved during normal development as confirmed by our PARP down-regulation studies. Constitutive PARP down-regulation resulted in blocked development while no effect was observed on D. discoideum growth. Interestingly, stage specific PARP down-regulation arrested development at the slug stage. Conclusion These results emphasize that PARP is essential for complex differentiation and its function may be linked to multicellularity. This is the first report where the involvement of PARP during normal multicellular development in D. discoideum, an ancient eukaryote, is established which could be of evolutionary significance. Thus our study adds one more role to the multitasking function of PARP.

Dicty_cDB: CFG822 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available 115581 |pid:none) Dictyostelium discoideum chromosom... 119 4e-26 BC077988_1( BC077988 |pid:none) Xenopus laevis achalasia...none) Homo sapiens mRNA for achalasia, a... 36 0.47 (Q9NRG9) RecName: Full=Aladin; AltName: Full=Adracalin;

Involvement of Sib Proteins in the Regulation of Cellular Adhesion in Dictyostelium discoideum▿ †

OpenAIRE

Cornillon, Sophie; Froquet, Romain; Cosson, Pierre

2008-01-01

Molecular mechanisms ensuring cellular adhesion have been studied in detail in Dictyostelium amoebae, but little is known about the regulation of cellular adhesion in these cells. Here, we show that cellular adhesion is regulated in Dictyostelium, notably by the concentration of a cellular secreted factor accumulating in the medium. This constitutes a quorum-sensing mechanism allowing coordinated regulation of cellular adhesion in a Dictyostelium population. In order to understand the mechani...

P2X receptor channels in endocrine glands

Czech Academy of Sciences Publication Activity Database

Stojilkovic, S. S.; Zemková, Hana

2013-01-01

Roč. 2, č. 4 (2013), s. 173-180 ISSN 2190-460X R&D Projects: GA ČR(CZ) GBP304/12/G069 Institutional support: RVO:67985823 Keywords : ATP * purinergic P2X receptor channels * pituitary * endocrine glands Subject RIV: ED - Physiology

Organometallic chemical vapor deposition and characterization of ZnGe(1-x)Si(x)P2-Ge alloys on GaP substrates

Science.gov (United States)

Xing, G. C.; Bachmann, Klaus J.; Posthill, J. B.; Timmons, M. L.

1993-01-01

The epitaxial growth of ZnGe(1-x)Si(x)P2-Ge alloys on GaP substrates by open tube organometallic chemical vapor deposition (OMCVD) is reported. The chemical composition of the alloys characterized by energy dispersive X-ray spectroscopy shows that alloys with x up to 0.13 can be deposited on (001) GaP. Epitaxial growth with mirror smooth surface morphology was achieved for x less than or equal to 0.05. Transmission electron microscopy (TEM) micrographs of these alloys show specular epitaxy and the absence of microstructural defects indicating a defect density of less than 10(exp 7) cm(sup -2). Selected area electron diffraction pattern of the alloy shows that the epitaxial layer crystallizes in the chalcopyrite structure with relatively weak superlattice reflections indicating certain degree of randomness in the cation sublattice. Hall measurements show that the alloys are p-type, like the unalloyed films; the carrier concentration, however, dropped about 10 times from 2 x 10(exp 18) to 2 x 10(exp 17) cm(sup -3). Absorption measurements indicate that the band tailing in the absorption spectra of the alloy was shifted about 0.04 eV towards shorter wavelength as compared to the unalloyed material. Diodes fabricated from the n(+)-GaP/p-ZnSiP2-ZnGeP2-Ge heterostructure at x = 0.05 have a reverse break-down voltage of -10.8 V and a reverse saturation current density of approximately 6 x 10(exp -8) A/sq cm.

Bismuth on copper (110): analysis of the c(2x2) and p(4x1) structures by surface x-ray diffraction

DEFF Research Database (Denmark)

Lottermoser, L.; Buslaps, T.; Johnson, R.L.

1997-01-01

Surface X-ray diffraction has been used to analyze the atomic structures of the Cu(110)-c(2 x 2)-Bi and Cu(110)-p(4 x 1)-Bi reconstructions with submonolayer coverages. A quasi-hexagonal c(2 x 2) adlayer structure is formed when half a monolayer of bismuth is deposited; the coverage corresponds...... to 1.08 x 10(-15) atoms cm(-2). There is one Bi atom per c(2 x 2) surface unit cell, and the nearest-neighbor distance on the planar overlayer was found to be 4.43 Angstrom. In the case of the p(4 x 1) reconstruction formed at a coverage of 0.75 monolayers, both the in-plane and out-of-plane data...

Mechanism of cAMP-induced H -efflux of Dictyostelium cells: a role ...

Indian Academy of Sciences (India)

Unknown

2.1 Culture conditions and induction of the development of D. discoideum. Strain Ax-2 was grown ..... the H+-entry pathway for passive leakage of protons. Thus, the formation of any .... Universal Academy Press) pp 3–13. Maeda Y, Inouye K ...

Dicty_cDB: SLD420 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available SL (Link to library) SLD420 (Link to dictyBase) - - - Contig-U16325-1 SLD420E (Link... to Original site) - - - - - - SLD420E 434 Show SLD420 Library SL (Link to library) Clone ID SLD420 (Link to dict...yBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U16325-1 Original site URL http://dict... 7 Homology vs DNA Score E Sequences producing significant alignments: (bits) Value N ( AF066071 ) Dict...yostelium discoideum SP85 (pspB) gene, comple... 860 0.0 1 ( AC117075 ) Dictyostelium

P2X7 Receptor Function in Bone-Related Cancer

Directory of Open Access Journals (Sweden)

Elena Adinolfi

2012-01-01

Full Text Available Modulation of tumor microenvironment by different mediators is central in determining neoplastic formation and progression. Among these molecules extracellular ATP is emerging as a good candidate in promoting cell growth, neovascularization, tumor-host interactions, and metastatization. This paper summarizes recent findings on expression and function of P2X7 receptor for extracellular ATP in primary and metastatic bone cancers. Search of mRNA expression microchip databases and literature analysis demonstrate a high expression of P2X7 in primary bone tumors as well as in other malignancies such as multiple myeloma, neuroblastoma, breast, and prostate cancer. Evidence that P2X7 triggers NFATc1, PI3K/Akt, ROCK, and VEGF pathways in osteoblasts promoting either primary tumor development or osteoblastic lesions is also reported. Moreover, P2X7 receptor is involved in osteoclast differentiation, RANKL expression, matrix metalloproteases and cathepsin secretion thus promoting bone resorption and osteolytic lesions. Taken together these data point to a pivotal role for the P2X7 receptor in bone cancer biology.

Knocking out P2X receptors reduces transmitter secretion in taste buds

Science.gov (United States)

Huang, Yijen A.; Stone, Leslie M.; Pereira, Elizabeth; Yang, Ruibiao; Kinnamon, John C.; Dvoryanchikov, Gennady; Chaudhari, Nirupa; Finger, Thomas E.; Kinnamon, Sue C.; Roper, Stephen D.

2011-01-01

In response to gustatory stimulation, taste bud cells release a transmitter, ATP, that activates P2X2 and P2X3 receptors on gustatory afferent fibers. Taste behavior and gustatory neural responses are largely abolished in mice lacking P2X2 and P2X3 receptors (P2X2 and P2X3 double knockout, or “DKO” mice). The assumption has been that eliminating P2X2 and P2X3 receptors only removes postsynaptic targets but that transmitter secretion in mice is normal. Using functional imaging, ATP biosensor cells, and a cell-free assay for ATP, we tested this assumption. Surprisingly, although gustatory stimulation mobilizes Ca2+ in taste Receptor (Type II) cells from DKO mice, as from wild type (WT) mice, taste cells from DKO mice fail to release ATP when stimulated with tastants. ATP release could be elicited by depolarizing DKO Receptor cells with KCl, suggesting that ATP-release machinery remains functional in DKO taste buds. To explore the difference in ATP release across genotypes, we employed reverse transcriptase (RT)-PCR, immunostaining, and histochemistry for key proteins underlying ATP secretion and degradation: Pannexin1, TRPM5, and NTPDase2 (ecto-ATPase) are indistinguishable between WT and DKO mice. The ultrastructure of contacts between taste cells and nerve fibers is also normal in the DKO mice. Finally, quantitative RT-PCR show that P2X4 and P2X7, potential modulators of ATP secretion, are similarly expressed in taste buds in WT and DKO taste buds. Importantly, we find that P2X2 is expressed in WT taste buds and appears to function as an autocrine, positive feedback signal to amplify taste-evoked ATP secretion. PMID:21940456

Knocking out P2X receptors reduces transmitter secretion in taste buds.

Science.gov (United States)

Huang, Yijen A; Stone, Leslie M; Pereira, Elizabeth; Yang, Ruibiao; Kinnamon, John C; Dvoryanchikov, Gennady; Chaudhari, Nirupa; Finger, Thomas E; Kinnamon, Sue C; Roper, Stephen D

2011-09-21

In response to gustatory stimulation, taste bud cells release a transmitter, ATP, that activates P2X2 and P2X3 receptors on gustatory afferent fibers. Taste behavior and gustatory neural responses are largely abolished in mice lacking P2X2 and P2X3 receptors [P2X2 and P2X3 double knock-out (DKO) mice]. The assumption has been that eliminating P2X2 and P2X3 receptors only removes postsynaptic targets but that transmitter secretion in mice is normal. Using functional imaging, ATP biosensor cells, and a cell-free assay for ATP, we tested this assumption. Surprisingly, although gustatory stimulation mobilizes Ca(2+) in taste Receptor (Type II) cells from DKO mice, as from wild-type (WT) mice, taste cells from DKO mice fail to release ATP when stimulated with tastants. ATP release could be elicited by depolarizing DKO Receptor cells with KCl, suggesting that ATP-release machinery remains functional in DKO taste buds. To explore the difference in ATP release across genotypes, we used reverse transcriptase (RT)-PCR, immunostaining, and histochemistry for key proteins underlying ATP secretion and degradation: Pannexin1, TRPM5, and NTPDase2 (ecto-ATPase) are indistinguishable between WT and DKO mice. The ultrastructure of contacts between taste cells and nerve fibers is also normal in the DKO mice. Finally, quantitative RT-PCR show that P2X4 and P2X7, potential modulators of ATP secretion, are similarly expressed in taste buds in WT and DKO taste buds. Importantly, we find that P2X2 is expressed in WT taste buds and appears to function as an autocrine, positive feedback signal to amplify taste-evoked ATP secretion.

Vesicular Trafficking Defects, Developmental Abnormalities, and Alterations in the Cellular Death Process Occur in Cell Lines that Over-Express Dictyostelium GTPase, Rab2, and Rab2 Mutants

Directory of Open Access Journals (Sweden)

Katherine Maringer

2014-08-01

Full Text Available Small molecular weight GTPase Rab2 has been shown to be a resident of pre-Golgi intermediates and required for protein transport from the ER to the Golgi complex, however, the function of Rab2 in Dictyostelium has yet to be fully characterized. Using cell lines that over-express DdRab2, as well as cell lines over-expressing constitutively active (CA, and dominant negative (DN forms of the GTPase, we report a functional role in vesicular transport specifically phagocytosis, and endocytosis. Furthermore, Rab2 like other GTPases cycles between an active GTP-bound and an inactive GDP-bound state. We found that this GTP/GDP cycle for DdRab2 is crucial for normal Dictyostelium development and cell–cell adhesion. Similar to Rab5 and Rab7 in C. elegans, we found that DdRab2 plays a role in programmed cell death, possibly in the phagocytic removal of apoptotic corpses.

P2X7 on mouse T cells: one channel, many functions

Directory of Open Access Journals (Sweden)

Björn eRissiek

2015-05-01

Full Text Available The P2X7 receptor is an adenosine triphosphate (ATP-gated cation channel that is expressed by several cells of the immune system. P2X7 is best known for its proinflammatory role in promoting inflammasome formation and release of mature IL-1β by innate immune cells. Mounting evidence indicates that P2X7 is also an important regulatory receptor of murine and human T cell functions. Murine T cells express a sensitive splice variant of P2X7 that can be activated either by non-covalent binding of ATP or, in the presence of nicotinamide adenine dinucleotide (NAD+, by its covalent ADP-ribosylation catalyzed by the ecto-ADP-ribosyltransferase ARTC2.2. Prolonged activation of P2X7 by either one of these pathways triggers the induction of T cell death. Conversely, lower concentrations of ATP can activate P2X7 to enhance T cell proliferation and production of IL-2. In this review we will highlight the molecular and cellular consequences of P2X7 activation on mouse T cells and its versatile role in T cell homeostasis and activation. Further, we will discuss important differences in the function of P2X7 on human and murine T cells.

Dicty_cDB: SSF210 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available SS (Link to library) SSF210 (Link to dictyBase) - - - Contig-U16581-1 SSF210P (Link... to Original site) SSF210F 183 SSF210Z 197 SSF210P 380 - - Show SSF210 Library SS (Link to library) Clone ID SSF210 (Link to dict...yBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U16581-1 Original site URL http://dict...nt alignments: (bits) Value N D16417 |D16417.1 Dictyostelium discoideum mRNA. 339...01670 Skin cDNA library Ictalurus punctatus cDNA 5' similar to Ictacalcin, mRNA s

BTG interacts with retinoblastoma to control cell fate in Dictyostelium.

Directory of Open Access Journals (Sweden)

Daniele Conte

Full Text Available BACKGROUND: In the genesis of many tissues, a phase of cell proliferation is followed by cell cycle exit and terminal differentiation. The latter two processes overlap: genes involved in the cessation of growth may also be important in triggering differentiation. Though conceptually distinct, they are often causally related and functional interactions between the cell cycle machinery and cell fate control networks are fundamental to coordinate growth and differentiation. A switch from proliferation to differentiation may also be important in the life cycle of single-celled organisms, and genes which arose as regulators of microbial differentiation may be conserved in higher organisms. Studies in microorganisms may thus contribute to understanding the molecular links between cell cycle machinery and the determination of cell fate choice networks. METHODOLOGY/PRINCIPAL FINDINGS: Here we show that in the amoebozoan D. discoideum, an ortholog of the metazoan antiproliferative gene btg controls cell fate, and that this function is dependent on the presence of a second tumor suppressor ortholog, the retinoblastoma-like gene product. Specifically, we find that btg-overexpressing cells preferentially adopt a stalk cell (and, more particularly, an Anterior-Like Cell fate. No btg-dependent preference for ALC fate is observed in cells in which the retinoblastoma-like gene has been genetically inactivated. Dictyostelium btg is the only example of non-metazoan member of the BTG family characterized so far, suggesting that a genetic interaction between btg and Rb predated the divergence between dictyostelids and metazoa. CONCLUSIONS/SIGNIFICANCE: While the requirement for retinoblastoma function for BTG antiproliferative activity in metazoans is known, an interaction of these genes in the control of cell fate has not been previously documented. Involvement of a single pathway in the control of mutually exclusive processes may have relevant implication in the

Statins and ATP regulate nuclear pAkt via the P2X7 purinergic receptor in epithelial cells

International Nuclear Information System (INIS)

Mistafa, Oras; Hoegberg, Johan; Stenius, Ulla

2008-01-01

Many studies have documented P2X7 receptor functions in cells of mesenchymal origin. P2X7 is also expressed in epithelial cells and its role in these cells remains largely unknown. Our data indicate that P2X7 regulate nuclear pAkt in epithelial cells. We show that low concentration of atorvastatin, a drug inhibiting HMG-CoA reductase and cholesterol metabolism, or the natural agonist extracellular ATP rapidly decreased the level of insulin-induced phosphorylated Akt in the nucleus. This effect was seen within minutes and was inhibited by P2X7 inhibitors. Experiments employing P2X7 siRNA and HEK293 cells heterologously expressing P2X7 and in vivo experiments further supported an involvement of P2X7. These data indicate that extracellular ATP and statins via the P2X7 receptor modulate insulin-induced Akt signaling in epithelial cells

Statins and ATP regulate nuclear pAkt via the P2X7 purinergic receptor in epithelial cells

Energy Technology Data Exchange (ETDEWEB)

Mistafa, Oras; Hoegberg, Johan [Institute of Environmental Medicine, Karolinska Institutet, Box 210, 17177 Stockholm (Sweden); Stenius, Ulla [Institute of Environmental Medicine, Karolinska Institutet, Box 210, 17177 Stockholm (Sweden)

2008-01-04

Many studies have documented P2X7 receptor functions in cells of mesenchymal origin. P2X7 is also expressed in epithelial cells and its role in these cells remains largely unknown. Our data indicate that P2X7 regulate nuclear pAkt in epithelial cells. We show that low concentration of atorvastatin, a drug inhibiting HMG-CoA reductase and cholesterol metabolism, or the natural agonist extracellular ATP rapidly decreased the level of insulin-induced phosphorylated Akt in the nucleus. This effect was seen within minutes and was inhibited by P2X7 inhibitors. Experiments employing P2X7 siRNA and HEK293 cells heterologously expressing P2X7 and in vivo experiments further supported an involvement of P2X7. These data indicate that extracellular ATP and statins via the P2X7 receptor modulate insulin-induced Akt signaling in epithelial cells.

P2X receptors as targets for the treatment of status epilepticus

Science.gov (United States)

Henshall, David C.; Diaz-Hernandez, Miguel; Miras-Portugal, M. Teresa; Engel, Tobias

2013-01-01

Prolonged seizures are amongst the most common neurological emergencies. Status epilepticus is a state of continuous seizures that is life-threatening and prompt termination of status epilepticus is critical to protect the brain from permanent damage. Frontline treatment comprises parenteral administration of anticonvulsants such as lorazepam that facilitate γ-amino butyric acid (GABA) transmission. Because status epilepticus can become refractory to anticonvulsants in a significant proportion of patients, drugs which act on different neurotransmitter systems may represent potential adjunctive treatments. P2X receptors are a class of ligand-gated ion channel activated by ATP that contributes to neuro- and glio-transmission. P2X receptors are expressed by both neurons and glia in various brain regions, including the hippocampus. Electrophysiology, pharmacology and genetic studies suggest certain P2X receptors are activated during pathologic brain activity. Expression of several members of the family including P2X2, P2X4, and P2X7 receptors has been reported to be altered in the hippocampus following status epilepticus. Recent studies have shown that ligands of the P2X7 receptor can have potent effects on seizure severity during status epilepticus and mice lacking this receptor display altered seizures in response to chemoconvulsants. Antagonists of the P2X7 receptor also modulate neuronal death, microglial responses and neuroinflammatory signaling. Recent work also found altered neuronal injury and inflammation after status epilepticus in mice lacking the P2X4 receptor. In summary, members of the P2X receptor family may serve important roles in the pathophysiology of status epilepticus and represent novel targets for seizure control and neuroprotection. PMID:24324404

Blockade of human P2X7 receptor function with a monoclonal antibody.

Science.gov (United States)

Buell, G; Chessell, I P; Michel, A D; Collo, G; Salazzo, M; Herren, S; Gretener, D; Grahames, C; Kaur, R; Kosco-Vilbois, M H; Humphrey, P P

1998-11-15

A monoclonal antibody (MoAb) specific for the human P2X7 receptor was generated in mice. As assessed by flow cytometry, the MoAb labeled human blood-derived macrophage cells natively expressing P2X7 receptors and cells transfected with human P2X7 but not other P2X receptor types. The MoAb was used to immunoprecipitate the human P2X7 receptor protein, and in immunohistochemical studies on human lymphoid tissue, P2X7 receptor labeling was observed within discrete areas of the marginal zone of human tonsil sections. The antibody also acted as a selective antagonist of human P2X7 receptors in several functional studies. Thus, whole cell currents, elicited by the brief application of 2',3'-(4-benzoyl)-benzoyl-ATP in cells expressing human P2X7, were reduced in amplitude by the presence of the MoAb. Furthermore, preincubation of human monocytic THP-1 cells with the MoAb antagonized the ability of P2X7 agonists to induce the release of interleukin-1beta.

Dicty_cDB: SHF448 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available SH (Link to library) SHF448 (Link to dictyBase) - - - Contig-U11503-1 SHF448E (Link...) Clone ID SHF448 (Link to dictyBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U11503-1 Ori...quence qkkqfsl*iy*YMIRKSNNFSILFAIFLKIVFVVSAPLCPNSTILLNYNILTVYNSSEGC GFNNEPICTSLKD...mnvnlkimrigltqlxisyrfy Frame C: qkkqfsl*iy*YMIRKSNNFSILFAIFLKIVFVVSAPLCPNSTILLNYNILTVYNSSEGC GFNNEPICTSLKDAV...nts: (bits) Value N AC115680 |AC115680.3 Dictyostelium discoideum chromosome 2 map 4915084-5005461 strain AX

The role of P2X receptors in bone biology

DEFF Research Database (Denmark)

Jørgensen, N R; Syberg, S; Ellegaard, M

2015-01-01

receptors regulate bone metabolism and especially for the P2X7 receptor an impressive amount of evidence has now documented its expression in osteoblasts, osteoclasts, and osteocytes as well as important functional roles in proliferation, differentiation, and function of the cells of bone. Key evidence has...... come from studies on murine knockout models and from pharmacologic studies on cells and animals. More recently, the role of P2X receptors in human bone diseases has been documented. Loss-of-functions polymorphisms in the P2X7 receptorare associated with bone loss and increased fracture risk. Very...

A Simple Retroelement Based Knock-Down System in Dictyostelium: Further Insights into RNA Interference Mechanisms.

Science.gov (United States)

Friedrich, Michael; Meier, Doreen; Schuster, Isabelle; Nellen, Wolfgang

2015-01-01

We have previously shown that the most abundant Dictyostelium discoideum retroelement DIRS-1 is suppressed by RNAi mechanisms. Here we provide evidence that both inverted terminal repeats have strong promoter activity and that bidirectional expression apparently generates a substrate for Dicer. A cassette containing the inverted terminal repeats and a fragment of a gene of interest was sufficient to activate the RNAi response, resulting in the generation of ~21 nt siRNAs, a reduction of mRNA and protein expression of the respective endogene. Surprisingly, no transitivity was observed on the endogene. This was in contrast to previous observations, where endogenous siRNAs caused spreading on an artificial transgene. Knock-down was successful on seven target genes that we examined. In three cases a phenotypic analysis proved the efficiency of the approach. One of the target genes was apparently essential because no knock-out could be obtained; the RNAi mediated knock-down, however, resulted in a very slow growing culture indicating a still viable reduction of gene expression. ADVANTAGES OF THE DIRS-1–RNAI SYSTEM: The knock-down system required a short DNA fragment (~400 bp) of the target gene as an initial trigger. Further siRNAs were generated by RdRPs since we have shown some siRNAs with a 5'-triphosphate group. Extrachromosomal vectors facilitate the procedure and allowed for molecular and phenotypic analysis within one week. The system provides an efficient and rapid method to reduce protein levels including those of essential genes.

DESENSITIZATION PROPERTIES OF P2X3 RECEPTORS SHAPING PAIN SIGNALLING

Directory of Open Access Journals (Sweden)

Rashid eGiniatullin

2013-12-01

Full Text Available ATP-gated P2X3 receptors are mostly expressed by nociceptive sensory neurons and participate in transduction of pain signals. P2X3 receptors show a combination of fast desensitization onset and slow recovery. Moreover, even low nanomolar agonist concentrations unable to evoke a response, can induce desensitization via a phenomenon called ‘high affinity desensitization’. We have also observed that recovery from desensitization is agonist-specific and can range from seconds to minutes. The recovery process displays unusually high temperature dependence. Likewise, recycling of P2X3 receptors in peri-membrane regions shows unexpectedly large temperature sensitivity. By applying kinetic modeling, we have previously shown that desensitization characteristics of P2X3 receptor are best explained with a cyclic model of receptor operation involving three agonist molecules binding a single receptor and that desensitization is primarily developing from the open receptor state. Mutagenesis experiments suggested that desensitization depends on a certain conformation of the ATP binding pocket and on the structure of the transmembrane domains forming the ion pore. Further molecular determinants of desensitization have been identified by mutating the intracellular N- and C-termini of P2X3 receptor. Unlike other P2X receptors, the P2X3 subtype is facilitated by extracellular calcium that acts via specific sites in the ectodomain neighboring the ATP binding pocket. Thus, substitution of serine275 in this region (called ‘left flipper’ converts the natural facilitation induced by extracellular calcium to receptor inhibition. Given such their strategic location in nociceptive neurons and unique desensitization properties, P2X3 receptors represent an attractive target for development of new analgesic drugs via promotion of desensitization aimed at suppressing chronic pain.

Autonomous and non-autonomous traits mediate social cooperation ...

Indian Academy of Sciences (India)

2011-07-08

Jul 8, 2011 ... In the trishanku (triA−) mutant of the social amoeba Dictyostelium discoideum, aggregates are smaller than usual and the spore mass is located mid-way up the stalk, not at the apex. We have monitored aggregate territory size, spore allocation and fruiting body morphology in chimaeric groups of ...

Dicty_cDB: Contig-U14279-1 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available Dictyostelium discoideum cDNA clone:ddc30k04, 5' ... 200 6e-48 1 ( EW967860 ) LS_13_N05_T7 Headlice composite...tobia irritans 1st Instar Larvae H... 42 3.5 1 ( EW966307 ) SFHL_01_A10_T7 Headlice composite library with a

Non-nucleotide Agonists Triggering P2X7 Receptor Activation and Pore Formation

Directory of Open Access Journals (Sweden)

Francesco Di Virgilio

2018-02-01

Full Text Available The P2X7 receptor (P2X7R is a ligand-gated plasma membrane ion channel belonging to the P2X receptor subfamily activated by extracellular nucleotides. General consensus holds that the physiological (and maybe the only agonist is ATP. However, scattered evidence generated over the last several years suggests that ATP might not be the only agonist, especially at inflammatory sites. Solid data show that NAD+ covalently modifies the P2X7R of mouse T lymphocytes, thus lowering the ATP threshold for activation. Other structurally unrelated agents have been reported to activate the P2X7R via a poorly understood mechanism of action: (a the antibiotic polymyxin B, possibly a positive allosteric P2X7R modulator, (b the bactericidal peptide LL-37, (c the amyloidogenic β peptide, and (d serum amyloid A. Some agents, such as Alu-RNA, have been suggested to activate the P2X7R acting on the intracellular N- or C-terminal domains. Mode of P2X7R activation by these non-nucleotide ligands is as yet unknown; however, these observations raise the intriguing question of how these different non-nucleotide ligands may co-operate with ATP at inflammatory or tumor sites. New information obtained from the cloning and characterization of the P2X7R from exotic mammalian species (e.g., giant panda and data from recent patch-clamp studies are strongly accelerating our understanding of P2X7R mode of operation, and may provide hints to the mechanism of activation of P2X7R by non-nucleotide ligands.

P2X Receptors and Synaptic Plasticity

Czech Academy of Sciences Publication Activity Database

Pankratov, Y.; Lalo, U.; Krishtal, A.; Verkhratsky, Alexei

2009-01-01

Roč. 158, č. 1 (2009), s. 137-148 ISSN 0306-4522 Institutional research plan: CEZ:AV0Z50390512 Keywords : ATP * P2X receptors * synaptic plasticity Subject RIV: FH - Neurology Impact factor: 3.292, year: 2009

Dicty_cDB: Contig-U10467-1 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available ments: (bits) Value N ( BJ433957 ) Dictyostelium discoideum cDNA clone:ddv23p10, 3' ... 1084 0.0 2 ( EU868611 ) Human enterovirus...73 ) Xenopus tropicalis cDNA clone MGC:172876 IMAGE:76... 44 7.1 1 ( EF063152 ) Human enterovirus... 71 strain E2005125-TW polyprote... 44 7.1 1 ( DQ868521 ) Human enterovirus 71 isolate E2006...249-TW VP1 stru... 44 7.1 1 ( DQ846663 ) Human enterovirus 71 isolate NTU1551-TW-06 VP1 st... 44 7.1 1 ( DQ846662 ) Human enterovirus... 71 isolate NTU1482-TW-06 VP1 st... 44 7.1 1 ( DQ841970 ) Human enterovirus 71 isol

Loss of Cln3 impacts protein secretion in the social amoeba Dictyostelium.

Science.gov (United States)

Huber, Robert J

2017-07-01

Neuronal ceroid lipofuscinosis (NCL), also referred to as Batten disease, is the most common form of childhood neurodegeneration. Mutations in CLN3 cause the most prevalent subtype of the disease, which manifests during early childhood and is currently untreatable. The precise function of the CLN3 protein is still not known, which has inhibited the development of targeted therapies. In the social amoeba Dictyostelium discoideum, loss of the CLN3 homolog, Cln3, reduces adhesion during early development, which delays streaming and aggregation. The results of the present study indicate that this phenotype may be at least partly due to aberrant protein secretion in cln3 - cells. It is well-established that Cln3 localizes primarily to the contractile vacuole (CV) system in Dictyostelium, and to a lesser extent, compartments of the endocytic pathway. Intriguingly, the CV system has been linked to the secretion of proteins that do not contain a signal peptide for secretion (i.e., unconventional protein secretion). Proteins that do contain a signal peptide are secreted via a conventional mechanism involving the endoplasmic reticulum, transport through the Golgi, and secretion via vesicle release. In this study, Cln3 was observed to co-localize with the Golgi marker wheat germ agglutinin suggesting that Cln3 participates in both secretion mechanisms. Chimeras of wild-type (WT) and cln3 - cells displayed delayed streaming and aggregation, and interestingly, cln3 - cells starved in conditioned media (CM) harvested from starving WT cells showed near normal timing of streaming and aggregation suggesting aberrant protein secretion in Cln3-deficient cells. Based on these observations, LC-MS/MS was used to reveal the protein content of CM from starved cells (mass spectrometry data are available via ProteomeXchange with identifier PXD004897). A total of 450 proteins were detected in WT and cln3 - CM, of which 3 were absent in cln3 - CM. Moreover, 12 proteins that were present in

An Improved Method for P2X7R Antagonist Screening.

Directory of Open Access Journals (Sweden)

Rômulo José Soares-Bezerra

Full Text Available ATP physiologically activates the P2X7 receptor (P2X7R, a member of the P2X ionotropic receptor family. When activated by high concentrations of ATP (i.e., at inflammation sites, this receptor is capable of forming a pore that allows molecules of up to 900 Da to pass through. This receptor is upregulated in several diseases, particularly leukemia, rheumatoid arthritis and Alzheimer's disease. A selective antagonist of this receptor could be useful in the treatment of P2X7R activation-related diseases. In the present study, we have evaluated several parameters using in vitro protocols to validate a high-throughput screening (HTS method to identify P2X7R antagonists. We generated dose-response curves to determine the EC50 value of the known agonist ATP and the ICs50 values for the known antagonists Brilliant Blue G (BBG and oxidized ATP (OATP. The values obtained were consistent with those found in the literature (0.7 ± 0.07 mM, 1.3-2.6 μM and 173-285 μM for ATP, BBG and OATP, respectively [corrected].The Z-factor, an important statistical tool that can be used to validate the robustness and suitability of an HTS assay, was 0.635 for PI uptake and 0.867 for LY uptake. No inter-operator variation was observed, and the results obtained using our improved method were reproducible. Our data indicate that our assay is suitable for the selective and reliable evaluation of P2X7 activity in multiwell plates using spectrophotometry-based methodology. This method might improve the high-throughput screening of conventional chemical or natural product libraries for possible candidate P2X7R antagonist or agonist.

An evolutionarily significant unicellular strategy in response to starvation stress in Dictyostelium social amoebae [v1; ref status: indexed, http://f1000r.es/3hg

Directory of Open Access Journals (Sweden)

Darja Dubravcic

2014-06-01

Full Text Available The social amoeba Dictyostelium discoideum is widely studied for its multicellular development program as a response to starvation and constitutes a model of choice in microbial cooperation studies. Aggregates of up to 106 cells form fruiting bodies containing two cell types: (i dormant spores (~80% that can persist for months in the absence of nutrients, and (ii dead stalk cells (~20% that promote the dispersion of the spores towards nutrient-rich areas. It is often overlooked that not all cells aggregate upon starvation. Using a new quantitative approach based on time-lapse fluorescence microscopy and a low ratio of reporting cells, we have quantified this fraction of non-aggregating cells. In realistic starvation conditions, up to 15% of cells do not aggregate, which makes this third cell fate a significant component of the population-level response of social amoebae to starvation. Non-aggregating cells have an advantage over cells in aggregates since they resume growth earlier upon arrival of new nutrients, but have a shorter lifespan under prolonged starvation. We find that phenotypic heterogeneities linked to cell nutritional state bias the representation of cells in the aggregating vs. non-aggregating fractions, and thus regulate population partitioning. Next, we report that the fraction of non-aggregating cells depends on genetic factors that regulate the timing of starvation, signal sensing efficiency and aggregation efficiency. In addition, interactions between clones in mixtures of non-isogenic cells affect the partitioning of each clone into both fractions. We further test the evolutionary significance of the non-aggregating cell fraction. The partitioning of cells into aggregating and non-aggregating fractions is optimal in fluctuating environments with an unpredictable duration of starvation periods. D. discoideum thus constitutes a model system lying at the intersection of microbial cooperation and bet hedging, defining a new

Journal of Biosciences | Indian Academy of Sciences

Indian Academy of Sciences (India)

pp 157-166 Articles. Ammonia differentially suppresses the cAMP chemotaxis of anterior-like cells and prestalk cells in Dictyostelium discoideum · Ira N Feit Erika J Medynski Michael J Rothrock · More Details Abstract Fulltext PDF. A drop assay for chemotaxis to cAMP confirms that both anterior-like cells (ALC) and prestalk ...

Nociceptive transmission and modulation via P2X receptors in central pain syndrome.

Science.gov (United States)

Kuan, Yung-Hui; Shyu, Bai-Chuang

2016-05-26

Painful sensations are some of the most frequent complaints of patients who are admitted to local medical clinics. Persistent pain varies according to its causes, often resulting from local tissue damage or inflammation. Central somatosensory pathway lesions that are not adequately relieved can consequently cause central pain syndrome or central neuropathic pain. Research on the molecular mechanisms that underlie this pathogenesis is important for treating such pain. To date, evidence suggests the involvement of ion channels, including adenosine triphosphate (ATP)-gated cation channel P2X receptors, in central nervous system pain transmission and persistent modulation upon and following the occurrence of neuropathic pain. Several P2X receptor subtypes, including P2X2, P2X3, P2X4, and P2X7, have been shown to play diverse roles in the pathogenesis of central pain including the mediation of fast transmission in the peripheral nervous system and modulation of neuronal activity in the central nervous system. This review article highlights the role of the P2X family of ATP receptors in the pathogenesis of central neuropathic pain and pain transmission. We discuss basic research that may be translated to clinical application, suggesting that P2X receptors may be treatment targets for central pain syndrome.

Dicty_cDB: Contig-U05033-1 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available 35269 Global-Ocean-Sampling_GS-34-01-01-1... 48 0.13 2 ( EY497680 ) CBBP3693.rev CBBP Hirudo medicinalis her...maphrodit... 50 0.13 1 ( EY493998 ) CBBP19771.rev CBBP Hirudo medicinalis hermaph...rodi... 50 0.13 1 ( EY487045 ) CBBP15563.fwd CBBP Hirudo medicinalis hermaphrodi... 50 0.13 1 ( EY487044 ) C...BBP15563.rev CBBP Hirudo medicinalis hermaphrodi... 50 0.13 1 ( AC116305 ) Dictyostelium discoideum chromoso

Correlation between Urothelial Differentiation and Sensory Proteins P2X3, P2X5, TRPV1, and TRPV4 in Normal Urothelium and Papillary Carcinoma of Human Bladder

Directory of Open Access Journals (Sweden)

Igor Sterle

2014-01-01

Full Text Available Terminal differentiation of urothelium is a prerequisite for blood-urine barrier formation and enables normal sensory function of the urinary bladder. In this study, urothelial differentiation of normal human urothelium and of low and high grade papillary urothelial carcinomas was correlated with the expression and localization of purinergic receptors (P2X3, and P2X5 and transient receptor potential vanilloid channels (TRPV1, and TRPV4. Western blotting and immunofluorescence of uroplakins together with scanning electron microscopy of urothelial apical surface demonstrated terminal differentiation of normal urothelium, partial differentiation of low grade carcinoma, and poor differentiation of high grade carcinoma. P2X3 was expressed in normal urothelium as well as in low grade carcinoma and in both cases immunolabeling was stronger in the superficial cells. P2X3 expression decreased in high grade carcinoma. P2X5 expression was detected in normal urothelium and in high grade carcinoma, while in low grade carcinoma its expression was diminished. The expression of TRPV1 decreased in low grade and even more in high grade carcinoma when compared with normal urothelium, while TRPV4 expression was unchanged in all samples. Our results suggest that sensory proteins P2X3 and TRPV1 are in correlation with urothelial differentiation, while P2X5 and TRPV4 have unique expression patterns.

Dicty_cDB: SFJ801 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available SF (Link to library) SFJ801 (Link to dictyBase) - - - - SFJ801P (Link to Original s...ite) SFJ801F 633 SFJ801Z 345 SFJ801P 978 - - Show SFJ801 Library SF (Link to library) Clone ID SFJ801 (Link to dict...yBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig - Original site URL http://dictycdb.biol.ts...t alignments: (bits) Value N U36937 |U36937.1 Dictyostelium discoideum calreticulin mRNA, complete cds. 1021...IpTrk_27_j08 Trunk kidney cDNA library Ictalurus punctatus cDNA 5' similar to ER-resident chaperone calretic

Dicty_cDB: [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VF (Link to library) VFB612 (Link to dictyBase) - - - Contig-U16272-1 VFB612P (Link... to Original site) VFB612F 631 VFB612Z 640 VFB612P 1271 - - Show VFB612 Library VF (Link to library) Clone ID VFB612 (Link to dict...yBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U16272-1 Original site URL http://dict...es producing significant alignments: (bits) Value N L08391 |L08391.1 Dictyostelium discoideum ribosomal prot...gi 4.0 %: plasma membrane >> prediction for VFB612 is nuc 5' end seq. ID VFB612F

Dicty_cDB: [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VF (Link to library) VFB847 (Link to dictyBase) - - - Contig-U16272-1 VFB847P (Link... to Original site) VFB847F 528 VFB847Z 749 VFB847P 1277 - - Show VFB847 Library VF (Link to library) Clone ID VFB847 (Link to dict...yBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U16272-1 Original site URL http://dict...core E Sequences producing significant alignments: (bits) Value N L08391 |L08391.1 Dictyostelium discoideum ...ear 24.0 %: cytoplasmic 4.0 %: cytoskeletal 4.0 %: plasma membrane 4.0 %: endoplasmic reticulum >> predictio

Dicty_cDB: [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VF (Link to library) VFC255 (Link to dictyBase) - - - Contig-U16272-1 VFC255P (Link... to Original site) VFC255F 198 VFC255Z 525 VFC255P 723 - - Show VFC255 Library VF (Link to library) Clone ID VFC255 (Link to dict...yBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U16272-1 Original site URL http://dict...oducing significant alignments: (bits) Value N L08391 |L08391.1 Dictyostelium discoideum ribosomal protein (...ic 32.0 %: nuclear 4.0 %: Golgi 4.0 %: mitochondrial >> prediction for VFC255 is cyt 5' end seq. ID VFC255F

Dicty_cDB: [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VF (Link to library) VFB559 (Link to dictyBase) - - - Contig-U16272-1 VFB559P (Link... to Original site) VFB559F 307 VFB559Z 624 VFB559P 931 - - Show VFB559 Library VF (Link to library) Clone ID VFB559 (Link to dict...yBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U16272-1 Original site URL http://dict...alignments: (bits) Value N L08391 |L08391.1 Dictyostelium discoideum ribosomal pr...toplasmic 16.0 %: mitochondrial 4.0 %: nuclear >> prediction for VFB559 is cyt 5' end seq. ID VFB559F 5' end

Phospholipase Cδ regulates germination of Dictyostelium spores

NARCIS (Netherlands)

Dijken, Peter van; Haastert, Peter J.M. van

2001-01-01

Background: Many eukaryotes, including plants and fungi make spores that resist severe environmental stress. The micro-organism Dictyostelium contains a single phospholipase C gene (PLC); deletion of the gene has no effect on growth, cell movement and differentiation. In this report we show that PLC

Eukaryotic cell flattening

Science.gov (United States)

Bae, Albert; Westendorf, Christian; Erlenkamper, Christoph; Galland, Edouard; Franck, Carl; Bodenschatz, Eberhard; Beta, Carsten

2010-03-01

Eukaryotic cell flattening is valuable for improving microscopic observations, ranging from bright field to total internal reflection fluorescence microscopy. In this talk, we will discuss traditional overlay techniques, and more modern, microfluidic based flattening, which provides a greater level of control. We demonstrate these techniques on the social amoebae Dictyostelium discoideum, comparing the advantages and disadvantages of each method.

Mutant p97 exhibits species-specific changes of its ATPase activity and compromises the UBXD9-mediated monomerisation of p97 hexamers.

Science.gov (United States)

Rijal, Ramesh; Arhzaouy, Khalid; Strucksberg, Karl-Heinz; Cross, Megan; Hofmann, Andreas; Schröder, Rolf; Clemen, Christoph S; Eichinger, Ludwig

2016-01-01

p97 (VCP) is a homo-hexameric triple-A ATPase that exerts a plethora of cellular processes. Heterozygous missense mutations of p97 cause at least five human neurodegenerative disorders. However, the specific molecular consequences of p97 mutations are hitherto widely unknown. Our in silico structural models of human and Dictyostelium p97 showed that the disease-causing human R93C, R155H, and R155C as well as Dictyostelium R154C, E219K, R154C/E219K p97 mutations constitute variations in surface-exposed locations. In-gel ATPase activity measurements of p97 monomers and hexamers revealed significant mutation- and species-specific differences. While all human p97 mutations led to an increase in ATPase activity, no changes could be detected for the Dictyostelium R154C mutant, which is orthologous to human R155C. The E219K mutation led to an almost complete loss of activity, which was partially recuperated in the R154C/E219K double-mutant indicating p97 inter-domain communication. By means of co-immunoprecipitation experiments we identified an UBX-domain containing Dictyostelium protein as a novel p97 interaction partner. We categorized all UBX-domain containing Dictyostelium proteins and named the interaction partner UBXD9. Pull-down assays and surface plasmon resonance analyses of Dictyostelium UBXD9 or the human orthologue TUG/ASPL/UBXD9 demonstrated direct interactions with p97 as well as species-, mutation- and ATP-dependent differences in the binding affinities. Sucrose density gradient assays revealed that both human and Dictyostelium UBXD9 proteins very efficiently disassembled wild-type, but to a lesser extent mutant p97 hexamers into monomers. Our results are consistent with a scenario in which p97 point mutations lead to differences in enzymatic activities and molecular interactions, which in the long-term result in a late-onset and progressive multisystem disease. Copyright © 2016 The Authors. Published by Elsevier GmbH.. All rights reserved.

Cleavage sites in the polypeptide precursors of poliovirus protein P2-X

International Nuclear Information System (INIS)

Selmer, B.L.; Hanecak, R.; Anderson, C.W.; Wimmer, E.

1981-01-01

Partial amino-terminal sequence analysis has been performed on the three major polypeptide products (P2-3b, P2-5b, and P2-X) from the central region (P2) of the poliovirus polyprotein, and this analysis precisely locates the amino termini of these products with respect to the nucleotide sequence of the poliovirus RNA genome. Like most of the products of the replicase region (P3), the amino termini of P2-5b and P2-X are generated by cleavage between glutamine and glycine residues. Thus, P2-5b and P2-X are probably both produced by the action of a singly (virus-encoded.) proteinase. The amino terminus of P2-3b, on the other hand, is produced by a cleavage between the carboxy-terminal tyrosine of VP1 and the glycine encoded by nucleotides 3381-3383. This result may suggest that more than one proteolytic activity is required for the complete processing of the poliovirus polyprotein

Dispatch. Dictyostelium chemotaxis: fascism through the back door?

Science.gov (United States)

Insall, Robert

2003-04-29

Aggregating Dictyostelium cells secrete cyclic AMP to attract their neighbours by chemotaxis. It has now been shown that adenylyl cyclase is enriched in the rear of cells, and this localisation is required for normal aggregation.

The P2X7 ATP receptor modulates renal cyst development in vitro

International Nuclear Information System (INIS)

Hillman, Kate A.; Woolf, Adrian S.; Johnson, Tanya M.; Wade, Angela; Unwin, Robert J.; Winyard, Paul J.D.

2004-01-01

P2X 7 , a piercing receptor, is expressed in renal collecting ducts as they undergo fulminant cysto genesis in the cpk/cpk mouse model of autosomal recessive polycystic kidney disease (ARPKD). Dissociated cpk/cpk kidneys generate cysts from cell aggregates within 24 h of suspension culture and we demonstrate that BzATP, a P2X 7 agonist, reduces cystogenesis. This effect is P2X 7 -specific, because: (i) equimolar concentrations of other purinergic agonists, ATP and UTP, had lesser effects and (ii) the P2X 7 inhibitor, oxidized ATP, abrogated the BzATP-mediated reduction in cystogenesis. BzATP did not significantly affect total cell number, proliferation, LDH release or caspase 3 activity, and zVAD-fmk, a caspase blocker, failed to modulate BzATP effects. In addition, this P2X 7 agonist did not significantly alter cyst size, probably excluding altered vectorial transport. In vivo, ATP was detected in cyst fluid from cpk/cpk kidneys; moreover, P2X 7 protein was also upregulated in human fetal ARPKD epithelia versus normal fetal collecting ducts. Thus, ATP may inhibit pathological renal cyst growth through P2X 7 signaling

The scavenger activity of the human P2X7 receptor differs from P2X7 pore function by insensitivity to antagonists, genetic variation and sodium concentration: Relevance to inflammatory brain diseases.

Science.gov (United States)

Ou, Amber; Gu, Ben J; Wiley, James S

2018-04-01

Activation of P2X7 receptors is widely recognised to initiate proinflammatory responses. However P2X7 also has a dual function as a scavenger receptor which is active in the absence of ATP and plasma proteins and may be important in central nervous system (CNS) diseases. Here, we investigated both P2X7 pore formation and its phagocytic function in fresh human monocytes (as a model of microglia) by measuring ATP-induced ethidium dye uptake and fluorescent bead uptake respectively. This was studied in monocytes expressing various polymorphic variants as well as in the presence of different P2X7 antagonists and ionic media. P2X7-mediated phagocytosis was found to account for about half of Latrunculin (or Cytochalasin D)-sensitive bead engulfment by fresh human monocytes. Monocytes harbouring P2X7 Ala348Thr or Glu496Ala polymorphic variants showed increase or loss of ethidium uptake respectively, but these changes in pore formation did not always correspond to the changes in phagocytosis of YG beads. Unlike pore function, P2X7-mediated phagocytosis was not affected by three potent selective P2X7 antagonists and remained identical in Na + and K + media. Taken together, our results show that P2X7 is a scavenger receptor with important function in the CNS but its phagocytic function has features distinct from its pore function. Both P2X7 pore formation and P2X7-mediated phagocytosis should be considered in the design of new P2X7 antagonists for the treatment of CNS diseases. Copyright © 2018 Elsevier B.V. All rights reserved.

Electronic structure and X-ray spectroscopic properties of YbNi_2P_2

International Nuclear Information System (INIS)

Shcherba, I.D.; Bekenov, L.V.; Antonov, V.N.; Noga, H.; Uskokovic, D.; Zhak, O.; Kovalska, M.V.

2016-01-01

Highlights: • We present new experimental and theoretical data for YbNi_2P_2. • The presence of divalent and trivalent Yb ion found in YbNi_2P_2. • The calculation show good agreement with the experimental measurements. - Abstract: X-ray absorption spectrum at the Yb L_3 edge and X-ray emission spectra of Ni and P at the K and L_2_,_3 edges have been studied experimentally and theoretically in the mixed valent compound YbNi_2P_2 with ThCr_2Si_2 type crystal structure. The electronic structure of YbNi_2P_2 is investigated using the fully relativistic Dirac linear muffin-tin orbital (LMTO) band-structure method. The effect of the spin–orbit (SO) interaction and Coulomb repulsion U on the electronic structure of YbNi_2P_2 is examined in the frame of the LSDA + SO + U method. The core-hole effect in the final states as well as the effect of the electric quadrupole E_2 transitions have been investigated. A good agreement between the theory and the experiment was found. Both the trivalent and the divalent Yb ions in YbNi_2P_2 are reflected in the experimentally measured Yb L_3 X-ray absorption spectrum simultaneously. We found that the best agreement between the experimental spectrum and sum of the theoretically calculated Yb"2"+ and Yb"3"+ spectra is achieved with 73% ytterbium ions in 2+ state and 27% ions in 3+ state.

P2X receptor-mediated ATP purinergic signaling in health and disease

Directory of Open Access Journals (Sweden)

Jiang LH

2012-09-01

Full Text Available Lin-Hua JiangSchool of Biomedical Sciences, Faculty of Biological Sciences, University of Leeds, Leeds, United KingdomAbstract: Purinergic P2X receptors are plasma membrane proteins present in a wide range of mammalian cells where they act as a cellular sensor, enabling cells to detect and respond to extracellular adenosine triphosphate (ATP, an important signaling molecule. P2X receptors function as ligand-gated Ca2+-permeable cationic channels that open upon ATP binding to elevate intracellular Ca2+ concentrations and cause membrane depolarization. In response to sustained activation, P2X receptors induce formation of a pore permeable to large molecules. P2X receptors also interact with distinct functional proteins and membrane lipids to form specialized signaling complexes. Studies have provided compelling evidence to show that such P2X receptor-mediated ATP-signaling mechanisms determine and regulate a growing number and diversity of important physiological processes, including neurotransmission, muscle contraction, and cytokine release. There is accumulating evidence to support strong causative relationships of altered receptor expression and function with chronic pain, inflammatory diseases, cancers, and other pathologies or diseases. Numerous high throughput screening drug discovery programs and preclinical studies have thus far demonstrated the proof of concepts that the P2X receptors are druggable targets and selective receptor antagonism is a promising therapeutics approach. This review will discuss the recent progress in understanding the mammalian P2X receptors with respect to the ATP-signaling mechanisms, physiological and pathophysiological roles, and development and preclinical studies of receptor antagonists.Keywords: extracellular ATP, ion channel, large pore, signaling complex, chronic pain, inflammatory diseases

Impaired P2X1 Receptor-Mediated Adhesion in Eosinophils from Asthmatic Patients.

Science.gov (United States)

Wright, Adam; Mahaut-Smith, Martyn; Symon, Fiona; Sylvius, Nicolas; Ran, Shaun; Bafadhel, Mona; Muessel, Michelle; Bradding, Peter; Wardlaw, Andrew; Vial, Catherine

2016-06-15

Eosinophils play an important role in the pathogenesis of asthma and can be activated by extracellular nucleotides released following cell damage or inflammation. For example, increased ATP concentrations were reported in bronchoalveolar lavage fluids of asthmatic patients. Although eosinophils are known to express several subtypes of P2 receptors for extracellular nucleotides, their function and contribution to asthma remain unclear. In this article, we show that transcripts for P2X1, P2X4, and P2X5 receptors were expressed in healthy and asthmatic eosinophils. The P2X receptor agonist α,β-methylene ATP (α,β-meATP; 10 μM) evoked rapidly activating and desensitizing inward currents (peak 18 ± 3 pA/pF at -60 mV) in healthy eosinophils, typical of P2X1 homomeric receptors, which were abolished by the selective P2X1 antagonist NF449 (1 μM) (3 ± 2 pA/pF). α,β-meATP-evoked currents were smaller in eosinophils from asthmatic patients (8 ± 2 versus 27 ± 5 pA/pF for healthy) but were enhanced following treatment with a high concentration of the nucleotidase apyrase (17 ± 5 pA/pF for 10 IU/ml and 11 ± 3 pA/pF for 0.32 IU/ml), indicating that the channels are partially desensitized by extracellular nucleotides. α,β-meATP (10 μM) increased the expression of CD11b activated form in eosinophils from healthy, but not asthmatic, donors (143 ± 21% and 108 ± 11% of control response, respectively). Furthermore, α,β-meATP increased healthy (18 ± 2% compared with control 10 ± 1%) but not asthmatic (13 ± 1% versus 10 ± 0% for control) eosinophil adhesion. Healthy human eosinophils express functional P2X1 receptors whose activation leads to eosinophil αMβ2 integrin-dependent adhesion. P2X1 responses are constitutively reduced in asthmatic compared with healthy eosinophils, probably as the result of an increase in extracellular nucleotide concentration. Copyright © 2016 by The American Association of Immunologists, Inc.

Dicty_cDB: SLC433 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available SL (Link to library) SLC433 (Link to dictyBase) - - - Contig-U16397-1 SLC433Z (Link... to Original site) - - SLC433Z 613 - - - - Show SLC433 Library SL (Link to library) Clone ID SLC433 (Link to...ycdb.biol.tsukuba.ac.jp/CSM/SL/SLC4-B/SLC433Q.Seq.d/ Representative seq. ID SLC43...3Z (Link to Original site) Representative DNA sequence >SLC433 (SLC433Q) /CSM/SL/SLC4-B/SLC433Q.Seq.d/ XXXXX...tyostelium discoideum slug cDNA, clone SLH872. 1134 0.0 1 ( AU034279 ) Dictyostelium discoideum slug cDNA, clone SLC4

Targeting of the P2X7 receptor in pancreatic cancer and stellate cells

DEFF Research Database (Denmark)

Giannuzzo, Andrea; Saccomano, Mara; Napp, Joanna

2016-01-01

of PDAC. In the in vitro studies we show that human PDAC cells with luciferase gene (PancTu-1 Luc cells) express high levels of P2X7R protein. Allosteric P2X7R antagonist AZ10606120 inhibited cell proliferation in basal conditions, indicating that P2X7R was tonically active. Extracellular ATP and Bz......ATP, to which the P2X7R is more sensitive, further affected cell survival and confirmed complex functionality of P2X7R. PancTu-1 Luc migration and invasion was reduced by AZ10606120, and it was stimulated by PSCs, but not by PSCs from P2X7(-/-) animals. PancTu-1 Luc cells were orthotopically transplanted...

Pleckstrin Homology Domain Diffusion in Dictyostelium Cytoplasm Studied Using Fluorescence Correlation Spectroscopy

NARCIS (Netherlands)

Engel, Ruchira; Hink, Mark A.; Bosgraaf, Leonard; Haastert, Peter J.M. van; Visser, Antonie J.W.G.

2004-01-01

The translocation of pleckstrin homology (PH) domain-containing proteins from the cytoplasm to the plasma membrane plays an important role in the chemotaxis mechanism of Dictyostelium cells. The diffusion of three PH domain-green fluorescent protein (GFP) fusions (PH2-GFP, PH10-GFP, and PH-CRAC

Pleckstrin homology domain diffusion in Dictyostelium cytoplasm studied using fluorescence correlation spectroscopy

NARCIS (Netherlands)

Ruchira, A.; Hink, M.A.; Bosgraaf, L.; Haastert, van P.J.M.; Visser, A.J.W.G.

2004-01-01

The translocation of pleckstrin homology (PH) domain-containing proteins from the cytoplasm to the plasma membrane plays an important role in the chemotaxis mechanism of Dictyostelium cells. The diffusion of three PH domain-green fluorescent protein (GFP) fusions (PH2-GFP, PH10-GFP, and PH-CRAC

Dicty_cDB: Contig-U05312-1 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available 246 ) PDUts1124F05 Porcine testis cDNA library I Sus sc... 48 0.32 1 ( CT631096 ) Danio rerio EST, clone ZF_mu... 46 1.2 1 ( CV877151 ) PDUts1160G12 Porcine testis cDNA library I Sus sc... 46 1.2 1 ( CT729188 ) Danio rerio EST, clone ZF_mu... 44 4.9 1 ( CV865498 ) PDUts1018G06 Porcine testis cDNA library I Sus sc... 44 4.9 1 ( CT735187 ) Danio rerio EST, clone ZF_mu...774433 ) McClintock41_B07.ab1 Homarus EST library project ... 54 0.005 1 ( AU269391 ) Dictyostelium discoideum vegetati...1 3'. 46 1.2 1 ( CK415565 ) AUF_IpPit_32_p21 Pituitary cDNA library Ictalurus...

P2X7 receptor-deficient mice are susceptible to bone cancer pain

DEFF Research Database (Denmark)

Hansen, Rikke Rie; Nielsen, Christian K.; Nasser, Arafat

2011-01-01

The purinergic P2X7 receptor is implicated in both neuropathic and inflammatory pain, and has been suggested as a possible target in pain treatment. However, the specific role of the P2X7 receptor in bone cancer pain is unknown. We demonstrated that BALB/cJ P2X7 receptor knockout (P2X7R KO) mice...... were susceptible to bone cancer pain and moreover had an earlier onset of pain-related behaviours compared with cancer-bearing, wild-type mice. Furthermore, acute treatment with the selective P2X7 receptor antagonist, A-438079, failed to alleviate pain-related behaviours in models of bone cancer pain...... with and without astrocyte activation (BALB/cJ or C3H mice inoculated with 4T1 mammary cancer cells or NCTC 2472 osteosarcoma cells, respectively), suggesting that astrocytic P2X7 receptors play a negligible role in bone cancer pain. The results support the hypothesis that bone cancer pain is a separate pain state...

Hyperglycemia-induced Renal P2X7 Receptor Activation Enhances Diabetes-related Injury

Directory of Open Access Journals (Sweden)

Robert I. Menzies

2017-05-01

Full Text Available Diabetes is a leading cause of renal disease. Glomerular mesangial expansion and fibrosis are hallmarks of diabetic nephropathy and this is thought to be promoted by infiltration of circulating macrophages. Monocyte chemoattractant protein-1 (MCP-1 has been shown to attract macrophages in kidney diseases. P2X7 receptors (P2X7R are highly expressed on macrophages and are essential components of pro-inflammatory signaling in multiple tissues. Here we show that in diabetic patients, renal P2X7R expression is associated with severe mesangial expansion, impaired glomerular filtration (≤40 ml/min/1.73 sq. m., and increased interstitial fibrosis. P2X7R activation enhanced the release of MCP-1 in human mesangial cells cultured under high glucose conditions. In mice, P2X7R-deficiency prevented glomerular macrophage attraction and collagen IV deposition; however, the more severe interstitial inflammation and fibrosis often seen in human diabetic kidney diseases was not modelled. Finally, we demonstrate that a P2X7R inhibitor (AZ11657312 can reduce renal macrophage accrual following the establishment of hyperglycemia in a model of diabetic nephropathy. Collectively these data suggest that P2X7R activation may contribute to the high prevalence of kidney disease found in diabetics.

P2X purinoceptors as a link between hyperexcitability and neuroinflammation in status epilepticus.

Science.gov (United States)

Henshall, David C; Engel, Tobias

2015-08-01

There remains a need for more efficacious treatments for status epilepticus. Prolonged seizures result in the release of ATP from cells which activates the P2 class of ionotropic and metabotropic purinoceptors. The P2X receptors gate depolarizing sodium and calcium entry and are expressed by both neurons and glia throughout the brain, and a number of subtypes are upregulated after status epilepticus. Recent studies have explored the in vivo effects of targeting ATP-gated P2X receptors in preclinical models of status epilepticus, with particular focus on the P2X7 receptor (P2X7R). The P2X7R mediates microglial activation and the release of the proepileptogenic inflammatory cytokine interleukin 1β. The receptor may also directly modulate neurotransmission and gliotransmission and promote the recruitment of immune cells into brain parenchyma. Data from our group and collaborators show that status epilepticus produced by intraamygdala microinjection of kainic acid increases P2X7R expression in the hippocampus and neocortex of mice. Antagonism of the P2X7R in the model reduced seizure severity, microglial activation and interleukin 1β release, and neuronal injury. Coadministration of a P2X7R antagonist with a benzodiazepine also provided seizure suppression in a model of drug-refractory status epilepticus when either treatment alone was minimally effective. More recently, we showed that status epilepticus in immature rats is also reduced by P2X7R antagonism. Together, these findings suggest that P2X receptors may be novel targets for seizure control and interruption of neuroinflammation after status epilepticus. This article is part of a Special Issue entitled "Status Epilepticus". Copyright © 2015 Elsevier Inc. All rights reserved.

Dicty_cDB: [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VF (Link to library) VFB826 (Link to dictyBase) - - - Contig-U16584-1 VFB826F (Link... to Original site) VFB826F 430 - - - - - - Show VFB826 Library VF (Link to library) Clone ID VFB826 (Link to dict...yBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U16584-1 Original site URL http://dict... (bits) Value N AC115577 |AC115577.2 Dictyostelium discoideum chromosome 2 map 4657875-4914984 strain AX4, c...vesicles of secretory system 4.0 %: endoplasmic reticulum >> prediction for VFB82

Dicty_cDB: [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VF (Link to library) VFB644 (Link to dictyBase) - - - Contig-U10683-1 VFB644F (Link... to Original site) VFB644F 519 - - - - - - Show VFB644 Library VF (Link to library) Clone ID VFB644 (Link to dict...yBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U10683-1 Original site URL http://dict...7 9 AC116920 |AC116920.2 Dictyostelium discoideum chromosome 2 map 3879572-407176... %: nuclear 8.0 %: mitochondrial 4.0 %: cytoskeletal 4.0 %: vesicles of secretory system >> predict

Dicty_cDB: Contig-U06875-1 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available . 44 3.6 1 ( DW405755 ) EST000176 Trichophyton rubrum cDNA library Tricho... 44 3.6 1 ( AU269367 ) Dictyostelium discoideum vegetati...5aa06.g2 hhd Oryza coarctata genomic clone ... 46 0.91 1 ( EV115075 ) 0120387 Brassica napus Root library Brassic...ES Homo sapiens cDNA 5', mRNA ... 46 0.91 1 ( CF872366 ) tric002xo14.b11 T.reesei mycelial culture...4 3.6 1 ( ES282448 ) PQ037G01.XT7 non-sporulating culture of P. brassi... 44 3.6 1 ( EL772758 ) Plate_11b_G10 Hibernati...ng 13-lined squirrel brain... 44 3.6 1 ( EC618786 ) S_F11_a1_093.ab1 Rabbit heart cDNA library Or

P2X7 receptor activates extracellular signal-regulated kinases ERK1 and ERK2 independently of Ca2+ influx

DEFF Research Database (Denmark)

Amstrup, Jan; Novak, Ivana

2003-01-01

P2X7 nucleotide receptors modulate a spectrum of cellular events in various cells including epithelia, such as exocrine pancreas. Although the pharmacology and channel properties of the P2X7 receptors have been studied intensively, signal transduction pathways are relatively unknown. In this study...... we applied a heterologous expression system of rat P2X7 receptors in HEK-293 cells. We followed the receptor expression and function using the enhanced green fluorescent protein (EGFP) tag, activation of intracellular proteins and increases in cellular Ca2+. EGFP-P2X7 receptors localized...... to the plasma membrane, clusters within the membrane and intracellularly. Stimulation of P2X7 receptors in HEK-293 cells led to an activation of extracellular signal-regulated kinases ERK1 and ERK2 and this activation was seen after just 1 min of stimulation with ATP. Using C- and N-terminal P2X7-receptor...

P2X receptors in the cardiovascular system and their potential as therapeutic targets in disease.

Science.gov (United States)

Ralevic, Vera

2015-01-01

This review considers the expression and roles of P2X receptors in the cardiovascular system in health and disease and their potential as therapeutic targets. P2X receptors are ligand gated ion channels which are activated by the endogenous ligand ATP. They are formed from the assembly of three P2X subunit proteins from the complement of seven (P2X1-7), which can associate to form homomeric or heteromeric P2X receptors. The P2X1 receptor is widely expressed in the cardiovascular system, being located in the heart, in the smooth muscle of the majority of blood vessels and in platelets. P2X1 receptors expressed in blood vessels can be activated by ATP coreleased with noradrenaline as a sympathetic neurotransmitter, leading to smooth muscle depolarisation and contraction. There is evidence that the purinergic component of sympathetic neurotransmission is increased in hypertension, identifying P2X1 receptors as a possible therapeutic target in this disorder. P2X3 and P2X2/3 receptors are expressed on cardiac sympathetic neurones and may, through positive feedback of neuronal ATP at this prejunctional site, amplify sympathetic neurotransmission. Activation of P2X receptors expressed in the heart increases cardiac myocyte contractility, and an important role of the P2X4 receptor in this has been identified. Deletion of P2X4 receptors in the heart depresses contractile performance in models of heart failure, while overexpression of P2X4 receptors has been shown to be cardioprotective, thus P2X4 receptors may be therapeutic targets in the treatment of heart disease. P2X receptors have been identified on endothelial cells. Although immunoreactivity for all P2X1-7 receptor proteins has been shown on the endothelium, relatively little is known about their function, with the exception of the endothelial P2X4 receptor, which has been shown to mediate endothelium-dependent vasodilatation to ATP released during shear stress. The potential of P2X receptors as therapeutic targets

Dicty_cDB: Contig-U11651-1 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available 0 1 ( BJ435790 ) Dictyostelium discoideum cDNA clone:ddv28j10, 3' ... 355 2e-93 1 ( EY481033 ) CBBP11161.rev CBBP Hirudo medicina...lis hermaphrodi... 36 1.6 2 ( EY486444 ) CBBP15196.rev CBBP Hirudo medicinalis hermaphr...odi... 36 1.6 2 ( EY503165 ) CBBP6773.rev CBBP Hirudo medicinalis hermaphrodit...... 36 1.6 2 ( EY494245 ) CBBP19905.rev CBBP Hirudo medicinalis hermaphrodi... 36 1.6 2 ( EY504499 ) CBBP7545.rev CBBP Hirudo medicina...lis hermaphrodit... 36 1.6 2 ( EY503685 ) CBBP7067.rev CBBP Hirudo medicinalis hermap

Dicty_cDB: SSA564 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available SS (Link to library) SSA564 (Link to dictyBase) - - - - SSA564F (Link to Original s...ite) SSA564F 653 - - - - - - Show SSA564 Library SS (Link to library) Clone ID SSA564 (Link to dictyBase) Atlas ID - NBRP ID - dict...yBase ID - Link to Contig - Original site URL http://dictycdb.biol.tsukuba.ac.jp/CSM/...NNTLKPKQTTKGFNIGGQPGNPTN*l--- Frame C: tlkfvmplvmkictslvlvlkrstk*rklfmmenshqtldgl...bits) Value N M77492 |M77492.1 Dictyostelium discoideum glycoprotein phosphorylase 2 (glpD) gene, complete c

Dicty_cDB: AFH742 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available AC115581 |pid:none) Dictyostelium discoideum chromosom... 477 e-133 BC077988_1( BC077988 |pid:none) Xenopus laevis achalasia...lus 2 days neonate thymus... 84 3e-17 BC067671_1( BC067671 |pid:none) Danio rerio achalasia, adrenocorti... ... |pid:none) Homo sapiens mRNA for achalasia, a... 79 4e-16 (Q9NRG9) RecName: Full...0826 fis, cl... 79 5e-16 BC120418_1( BC120418 |pid:none) Bos taurus achalasia, adrenocortic... 80 6e-16 EF14

Dicty_cDB: VFH121 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VF (Link to library) VFH121 (Link to dictyBase) - - - - VFH121P (Link to Original s...ite) VFH121F 632 VFH121Z 144 VFH121P 776 - - Show VFH121 Library VF (Link to library) Clone ID VFH121 (Link to dict...yBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig - Original site URL http://dictycdb.biol.ts...nificant alignments: (bits) Value N U36937 |U36937.1 Dictyostelium discoideum calreticulin mRNA, complete cd...es cDNA clone hj81f04, mRNA sequence. 36 7e-04 3 CB937139 |CB937139.1 IpCGJx13_12_E07_23 IpCGJx13 Ictalurus

Dicty_cDB: [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VF (Link to library) VFB856 (Link to dictyBase) - - - Contig-U16605-1 VFB856P (Link... to Original site) VFB856F 552 VFB856Z 535 VFB856P 1087 - - Show VFB856 Library VF (Link to library) Clone ID VFB856 (Link to dict...yBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U16605-1 Original site URL http://dict...own update 2004.12.25 Homology vs DNA Score E Sequences producing significant alignments: (bits) Value N L36204 |L36204.1 Dict...ds. 922 0.0 6 U72746 |U72746.1 Dictyostelium discoideum cysteine proteinase (cprG) mRNA, complete cds. 131 e

Dicty_cDB: [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VF (Link to library) VFC241 (Link to dictyBase) - - - Contig-U16272-1 VFC241P (Link... to Original site) VFC241F 369 VFC241Z 514 VFC241P 883 - - Show VFC241 Library VF (Link to library) Clone ID VFC241 (Link to dict...yBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U16272-1 Original site URL http://dict...ments: (bits) Value N L08391 |L08391.1 Dictyostelium discoideum ribosomal protein (L3) gene, complete cds. 6...0 m3a: 0.00 m3b: 0.00 m_ : 1.00 68.0 %: nuclear 20.0 %: cytoplasmic 8.0 %: mitochondrial 4.0 %: peroxisomal >> predict

Dicty_cDB: [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VF (Link to library) VFC161 (Link to dictyBase) - - - Contig-U15456-1 VFC161P (Link... to Original site) VFC161F 564 VFC161Z 562 VFC161P 1126 - - Show VFC161 Library VF (Link to library) Clone ID VFC161 (Link to dict...yBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U15456-1 Original site URL http://dict...Value N Y16962 |Y16962.1 Dictyostelium discoideum mRNA for cathepsin D. 1025 0.0 5 AJ243946 |AJ243946.1 Dict...plasma membrane 4.0 %: vesicles of secretory system >> prediction for VFC161 is nuc 5' end seq. ID VFC161F 5

Rab5 regulates internalisation of P2X4 receptors and potentiation by ivermectin.

Science.gov (United States)

Stokes, Leanne

2013-03-01

The P2X4 receptor is an ATP-gated ion channel expressed in neurons, endothelia and immune cells. Plasma membrane expression of P2X4 is regulated by dynamin-dependent endocytosis, and this study identifies a Rab5-dependent pathway of receptor internalisation. Expression of Rab5 constructs altered the distribution of P2X4 in HEK-293 cells, and both constitutive internalisation and agonist-induced desensitisation of P2X4 were increased by co-expression of wild-type Rab5 or constitutively active Rab5 (Q79L). Expression of inactive dynamin K44A and Rab5 S34N constructs abolished agonist-induced desensitisation, suggesting internalisation as the underlying mechanism. Blocking P2X4 internalisation in this way also abolished potentiation of ATP-induced currents by the allosteric modulator ivermectin. This suggests that the dynamin-Rab5 internalisation pathway is essential for the ivermectin potentiation effect. In agreement with this hypothesis, the co-expression of wild-type dynamin, wild-type Rab5 or active Rab5 (Q79L) could increase the potentiation of the ATP-induced P2X4 response by ivermectin. These findings highlight Rab5 GTPase as a key regulator of P2X4 receptor cell surface expression and internalisation.

Novel protective role of endogenous cardiac myocyte P2X4 receptors in heart failure.

Science.gov (United States)

Yang, Tiehong; Shen, Jian-bing; Yang, Ronghua; Redden, John; Dodge-Kafka, Kimberly; Grady, James; Jacobson, Kenneth A; Liang, Bruce T

2014-05-01

Heart failure (HF), despite continuing progress, remains a leading cause of mortality and morbidity. P2X4 receptors (P2X4R) have emerged as potentially important molecules in regulating cardiac function and as potential targets for HF therapy. Transgenic P2X4R overexpression can protect against HF, but this does not explain the role of native cardiac P2X4R. Our goal is to define the physiological role of endogenous cardiac myocyte P2X4R under basal conditions and during HF induced by myocardial infarction or pressure overload. Mice established with conditional cardiac-specific P2X4R knockout were subjected to left anterior descending coronary artery ligation-induced postinfarct or transverse aorta constriction-induced pressure overload HF. Knockout cardiac myocytes did not show P2X4R by immunoblotting or by any response to the P2X4R-specific allosteric enhancer ivermectin. Knockout hearts showed normal basal cardiac function but depressed contractile performance in postinfarct and pressure overload models of HF by in vivo echocardiography and ex vivo isolated working heart parameters. P2X4R coimmunoprecipitated and colocalized with nitric oxide synthase 3 (eNOS) in wild-type cardiac myocytes. Mice with cardiac-specific P2X4R overexpression had increased S-nitrosylation, cyclic GMP, NO formation, and were protected from postinfarct and pressure overload HF. Inhibitor of eNOS, L-N(5)-(1-iminoethyl)ornithine hydrochloride, blocked the salutary effect of cardiac P2X4R overexpression in postinfarct and pressure overload HF as did eNOS knockout. This study establishes a new protective role for endogenous cardiac myocyte P2X4R in HF and is the first to demonstrate a physical interaction between the myocyte receptor and eNOS, a mediator of HF protection. © 2014 American Heart Association, Inc.

Characterization of two unusual guanylyl cyclases from Dictyostelium

NARCIS (Netherlands)

Roelofs, Jeroen; Haastert, Peter J.M. van

2002-01-01

Guanylyl cyclase A (GCA) and soluble guanylyl cyclase (sGC) encode GCs in Dictyostelium and have a topology similar to 12-transmembrane and soluble adenylyl cyclase, respectively. We demonstrate that all detectable GC activity is lost in a cell line in which both genes have been inactivated. Cell

Structural basis for subtype-specific inhibition of the P2X7 receptor

Energy Technology Data Exchange (ETDEWEB)

Karasawa, Akira; Kawate, Toshimitsu (Cornell)

2016-12-09

<p>The P2X7 receptor is a non-selective cation channel activated by extracellular adenosine triphosphate (ATP). Chronic activation of P2X7 underlies many health problems such as pathologic pain, yet we lack effective antagonists due to poorly understood mechanisms of inhibition. Here we present crystal structures of a mammalian P2X7 receptor complexed with five structurally-unrelated antagonists. Unexpectedly, these drugs all bind to an allosteric site distinct from the ATP-binding pocket in a groove formed between two neighboring subunits. This novel drug-binding pocket accommodates a diversity of small molecules mainly through hydrophobic interactions. Functional assays propose that these compounds allosterically prevent narrowing of the drug-binding pocket and the turret-like architecture during channel opening, which is consistent with a site of action distal to the ATP-binding pocket. These novel mechanistic insights will facilitate the development of P2X7-specific drugs for treating human diseases.p>

Dicty_cDB: [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VF (Link to library) VFB595 (Link to dictyBase) - - - Contig-U09552-1 VFB595E (Link...) Clone ID VFB595 (Link to dictyBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U09552-1 Ori...KLLKSDNWISTCQNLIQEYEPQ IIAVVEGFMAPSELCQKIKFCSSSSSTNDFDFIGSSTTDCEICTFISGYAENFLEENKTL EDIIKVVDDFCKILPAAYKTDCVA...A: VEGSGECLVCEFISEKIVTYLEANQTETQILQYLDNDCKLLKSDNWISTCQNLIQEYEPQ IIAVVEGFMAPSELCQKIKFCSSSSSTNDFDFIGSSTTDCEICT... Sequences producing significant alignments: (bits) Value N U66367 |U66367.1 Dictyostelium discoideum SapA (

Dicty_cDB: VHK273 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VH (Link to library) VHK273 (Link to dictyBase) - - - Contig-U16260-1 - (Link to Original site) VHK2...73F 620 - - - - - - Show VHK273 Library VH (Link to library) Clone ID VHK273 (Link to dicty...iol.tsukuba.ac.jp/CSM/VH/VHK2-D/VHK273Q.Seq.d/ Representative seq. ID - (Link to ...Original site) Representative DNA sequence >VHK273 (VHK273Q) /CSM/VH/VHK2-D/VHK273Q.Seq.d/ AACTCTCGAGTGCAAAA...27874 ) Dictyostelium discoideum cDNA clone:ddv63k23, 5' ... 1170 0.0 1 ( BJ42787

Dicty_cDB: VHK256 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VH (Link to library) VHK256 (Link to dictyBase) - - - Contig-U16260-1 - (Link to Original site) VHK2...56F 620 - - - - - - Show VHK256 Library VH (Link to library) Clone ID VHK256 (Link to dicty...iol.tsukuba.ac.jp/CSM/VH/VHK2-C/VHK256Q.Seq.d/ Representative seq. ID - (Link to ...Original site) Representative DNA sequence >VHK256 (VHK256Q) /CSM/VH/VHK2-C/VHK256Q.Seq.d/ AACTCTCGAGTGCAAAA...27874 ) Dictyostelium discoideum cDNA clone:ddv63k23, 5' ... 1170 0.0 1 ( BJ42787

Dicty_cDB: [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VF (Link to library) VFB703 (Link to dictyBase) - - - Contig-U14188-1 VFB703Z (Link... to Original site) - - VFB703Z 680 - - - - Show VFB703 Library VF (Link to library) Clone ID VFB703 (Link to dict...yBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U14188-1 Original site URL http://dict...ENCE, 18 unordered pieces. 36 0.038 5 AC115680 |AC115680.2 Dictyostelium discoideum chromosome 2 map 4415041...4-226K1, *** SEQUENCING IN PROGRESS ***, 52 unordered pieces. 46 0.31 5 AC116977 |AC116977.2 Dict

Dicty_cDB: [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VF (Link to library) VFC124 (Link to dictyBase) - - - Contig-U12017-1 VFC124Z (Link... to Original site) - - VFC124Z 496 - - - - Show VFC124 Library VF (Link to library) Clone ID VFC124 (Link to dict...yBase) Atlas ID - NBRP ID - dictyBase ID - Link to Contig Contig-U12017-1 Original site URL http://dict...e E Sequences producing significant alignments: (bits) Value N AC116957 |AC116957.2 Dictyostelium discoideum... chromosome 2 map 1685067-2090751 strain AX4, complete sequence. 835 0.0 2 U67089 |U67089.1 Dict

Dicty_cDB: SLH264 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available SL (Link to library) SLH264 (Link to dictyBase) - - - Contig-U16382-1 SLH264Z (Link... to Original site) - - SLH264Z 532 - - - - Show SLH264 Library SL (Link to library) Clone ID SLH264 (Link to...ycdb.biol.tsukuba.ac.jp/CSM/SL/SLH2-C/SLH264Q.Seq.d/ Representative seq. ID SLH26...4Z (Link to Original site) Representative DNA sequence >SLH264 (SLH264Q) /CSM/SL/SLH2-C/SLH264Q.Seq.d/ XXXXX...g significant alignments: (bits) Value N ( AU039244 ) Dictyostelium discoideum slug cDNA, clone SLH2

Symmetry-Breaking Transitions in RECuAs_2-xP_x (RE=Sm, Gd, Ho, and Er)

Energy Technology Data Exchange (ETDEWEB)

Mozharivskyj, Yurij [Iowa State Univ., Ames, IA (United States)

2002-01-01

Structural changes resulting in lower symmetries can be understood in terms of electronic instabilities and Coulomb interactions. The interplay of these two interrelated factors is complicated and difficult to analyze. The RECuAs_2-xP _x phases, because of the variation in the chemical content (As/P substitution), allow, with the aid of band structures, Madelung energies and Landau theory, a partial unraveling of the forces important in the symmetry-breaking transitions in RECuAs_2-xP _x (RE = Sm, Gd, Ho and Er). Distortions of the P layers in SmCu_1.15P₂, GdCuP_2.20 and ErCuP₂ are usefully thought of as generalized Peierls distortions, i.e., they lower the electronic (and total) energy and lead to more stable structures. On the other hand, the P4/nmm → Pmmn transitions, which are observed in all studied arsenophosphide series and occur upon substitution of P for As, originate from the B1g vibrational mode and are structural adaptations to smaller P atoms. These transitions provide tighter atomic packing and better Coulomb interactions. Configurational contribution to the entropy becomes important in stabilizing the mixed occupancy in the RECuAs _2-xP _x arsenophosphides. While geometric and electronic factors favor separation of the As and P atoms over two different crystallographic sites, configurational entropy stabilizes the As/P mixing on these two sites.;Progress in the research on RECuAs_2-xP _x was dependent upon the ability of Landau theory to predict, explain and dismiss structural models and transitions. The space group Pmmn (arising from the B 1g vibrational mode) in all mixed arsenophosphides and the existence of these mixed arsenophosphides followed from the analysis of GdCuAs ₂ and GdCuP₂, using Landau theory. The impossibility of obtaining the high-symmetry structure (P4/nmm) and the low symmetry structure

Role of P2X7 on steroid synthesis in murine luteal cells

Directory of Open Access Journals (Sweden)

Chunping Zhang

2016-03-01

Full Text Available The extracellular adenosine triphosphate (ATP regulates different cellular functions through activating purinergic receptors as a signalling molecule or neurotransmitter. P2X7 is highly expressed in murine small luteal cells. In this study, murine luteal cells were cultured in vitro and treated with P2X7 agonists – ATP and 2′(3′-O-(4-benzoyl-benzoyl-adenosine 50-triphosphate (BzATP and with P2X7 antagonist – brilliant blue G (BBG. We found that ATP and BzATP increased the production of progesterone and had no influence on the production of estradiol. BBG reversed the effect of BzATP and ATP. Further studies demonstrated that ATP and BzATP promoted the expression of CYP11A. These results revealed that P2X7 receptor activation is involved in the steroid synthesis in corpus luteum.

Prostaglandin E2 potentiation of P2X3 receptor mediated currents in dorsal root ganglion neurons

Directory of Open Access Journals (Sweden)

Huang Li-Yen

2007-08-01

Full Text Available Abstract Prostaglandin E2 (PGE2 is a well-known inflammatory mediator that enhances the excitability of DRG neurons. Homomeric P2X3 and heteromeric P2X2/3 receptors are abundantly expressed in dorsal root ganglia (DRG neurons and participate in the transmission of nociceptive signals. The interaction between PGE2 and P2X3 receptors has not been well delineated. We studied the actions of PGE2 on ATP-activated currents in dissociated DRG neurons under voltage-clamp conditions. PGE2 had no effects on P2X2/3 receptor-mediated responses, but significantly potentiated fast-inactivating ATP currents mediated by homomeric P2X3 receptors. PGE2 exerted its action by activating EP3 receptors. To study the mechanism underlying the action of PGE2, we found that the adenylyl cyclase activator, forskolin and the membrane-permeable cAMP analogue, 8-Br-cAMP increased ATP currents, mimicking the effect of PGE2. In addition, forskolin occluded the enhancement produced by PGE2. The protein kinase A (PKA inhibitors, H89 and PKA-I blocked the PGE2 effect. In contrast, the PKC inhibitor, bisindolymaleimide (Bis did not change the potentiating action of PGE2. We further showed that PGE2 enhanced α,β-meATP-induced allodynia and hyperalgesia and the enhancement was blocked by H89. These observations suggest that PGE2 binds to EP3 receptors, resulting in the activation of cAMP/PKA signaling pathway and leading to an enhancement of P2X3 homomeric receptor-mediated ATP responses in DRG neurons.

Dicty_cDB: [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VF (Link to library) VFB668 (Link to dictyBase) - G00394 DDB0168247 Contig-U09555-1...brary) Clone ID VFB668 (Link to dictyBase) Atlas ID - NBRP ID G00394 dictyBase ID DDB0168247 Link to Contig ...Contig-U09555-1 Original site URL http://dictycdb.biol.tsukuba.ac.jp/CSM/VF/VFB6-...uences producing significant alignments: (bits) Value N AC117072 |AC117072.2 Dictyostelium discoideum chromo...tein Score E Sequences producing significant alignments: (bits) Value AC117076_25

Dicty_cDB: [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available VF (Link to library) VFB677 (Link to dictyBase) - G02518 DDB0188526 Contig-U02066-1...brary) Clone ID VFB677 (Link to dictyBase) Atlas ID - NBRP ID G02518 dictyBase ID DDB0188526 Link to Contig ...Contig-U02066-1 Original site URL http://dictycdb.biol.tsukuba.ac.jp/CSM/VF/VFB6-...nfnytrgccsygdl*wy*gtr*tih*kdr*lwcassfenfndtictit mfnr**sstwfeilskrsih*rgikc*hd*nh..._25( AC116960 |pid:none) Dictyostelium discoideum chromoso... 90 2e-16 CP001276_520( CP001276 |pid:none) The