Sample records for vam3 vam7 vti1

  1. VAM3D-CG configuration management plan

    International Nuclear Information System (INIS)

    Langford, D.W.


    The VAM3D-CG computer code has been licensed for use at Hanford, from HydroGeologic, Inc., of Herndon, VA. Version 2.4b has been installed on the 3200GWW workstations, and is currently under configuration management. The purpose of this report is to describe the installation and configuration management of VAM3D-CG on the Hanford Computer System. VAM3D-CG is written in standard FORTRAN F77

  2. The SNARE protein vti1a functions in dense-core vesicle biogenesis

    DEFF Research Database (Denmark)

    Walter, Alexander M; Kurps, Julia; de Wit, Heidi


    overlapping with syntaxin-6. Exocytosis is impaired in vti1a null cells, partly due to fewer Ca(2+)-channels at the plasma membrane, partly due to fewer vesicles of reduced size and synaptobrevin-2 content. In contrast, release kinetics and Ca(2+)-sensitivity remain unchanged, indicating that the final fusion......The SNARE protein vti1a is proposed to drive fusion of intracellular organelles, but recent data also implicated vti1a in exocytosis. Here we show that vti1a is absent from mature secretory vesicles in adrenal chromaffin cells, but localizes to a compartment near the trans-Golgi network, partially...... reaction leading to transmitter release is unperturbed. Additional deletion of the closest related SNARE, vti1b, does not exacerbate the vti1a phenotype, and vti1b null cells show no secretion defects, indicating that vti1b does not participate in exocytosis. Long-term re-expression of vti1a (days...

  3. A Combined Experimental and Computational Study of Vam3, a Derivative of Resveratrol, and Syk Interaction

    Directory of Open Access Journals (Sweden)

    Ming Jiang


    Full Text Available Spleen tyrosine kinase (Syk plays an indispensable role through preliminary extracellular antigen-induced crosslinking of Fc receptor (FcR in the pathogenesis of autoimmune disorders, such as rheumatoid arthritis. In this study, we identify Vam3, a dimeric derivative of resveratrol isolated from grapes, as an ATP-competitive inhibitor of Syk with an IC50 of 62.95 nM in an in vitro kinase assay. Moreover, docking and molecular dynamics simulation approaches were performed to get more detailed information about the binding mode of Vam3 and Syk. The results show that 11b-OH on ring-C and 4b-OH on ring-D could form two hydrogen bonds with Glu449 and Phe382 of Syk, respectively. In addition, arene-cation interaction between ring-D of Vam3 and Lys402 of Syk was also observed. These results indicate that ring-C and D play an essential role in Vam3–Syk interaction. Our studies may be helpful in the structural optimization of Vam3, and also aid the design of novel Syk inhibitors in the future.

  4. DNA watermarks: A proof of concept

    Directory of Open Access Journals (Sweden)

    Barnekow Angelika


    Full Text Available Abstract Background DNA-based watermarks are helpful tools to identify the unauthorized use of genetically modified organisms (GMOs protected by patents. In silico analyses showed that in coding regions synonymous codons can be used to insert encrypted information into the genome of living organisms by using the DNA-Crypt algorithm. Results We integrated an authenticating watermark in the Vam7 sequence. For our investigations we used a mutant Saccharomyces cerevisiae strain, called CG783, which has an amber mutation within the Vam7 sequence. The CG783 cells are unable to sporulate and in addition display an abnormal vacuolar morphology. Transformation of CG783 with pRS314 Vam7 leads to a phenotype very similar to the wildtype yeast strain CG781. The integrated watermark did not influence the function of Vam7 and the resulting phenotype of the CG783 cells transformed with pRS314 Vam7-TB shows no significant differences compared to the CG783 cells transformed with pRS314 Vam7. Conclusion From our experiments we conclude that the DNA watermarks produced by DNA-Crypt do not influence the translation from mRNA into protein. By analyzing the vacuolar morphology, growth rate and ability to sporulate we confirmed that the resulting Vam7 protein was functionally active.

  5. Hydrological methods preferentially recover cesium from nuclear waste salt cake

    International Nuclear Information System (INIS)

    Brooke, J.N.; Hamm, L.L.


    The Savannah River Site is treating high level radioactive waste in the form of insoluble solids (sludge), crystallized salt (salt cake), and salt solutions. High costs and operational concerns have prompted DOE to look for ways to improve the salt cake treatment process. A numerical model was developed to evaluate the feasibility of pump and treat technology for extracting cesium from salt cake. A modified version of the VAM3DCG code was used to first establish a steady-state flow field, then to simulate 30 days of operation. Simulation results suggest that efficient cesium extraction can be obtained with low displacement volumes. The actual extraction process will probably be less impressive because of nonuniform properties. 2 refs., 2 figs

  6. Hanford statewide groundwater flow and transport model calibration report

    International Nuclear Information System (INIS)

    Law, A.; Panday, S.; Denslow, C.; Fecht, K.; Knepp, A.


    This report presents the results of the development and calibration of a three-dimensional, finite element model (VAM3DCG) for the unconfined groundwater flow system at the Hanford Site. This flow system is the largest radioactively contaminated groundwater system in the United States. Eleven groundwater plumes have been identified containing organics, inorganics, and radionuclides. Because groundwater from the unconfined groundwater system flows into the Columbia River, the development of a groundwater flow model is essential to the long-term management of these plumes. Cost effective decision making requires the capability to predict the effectiveness of various remediation approaches. Some of the alternatives available to remediate groundwater include: pumping contaminated water from the ground for treatment with reinjection or to other disposal facilities; containment of plumes by means of impermeable walls, physical barriers, and hydraulic control measures; and, in some cases, management of groundwater via planned recharge and withdrawals. Implementation of these methods requires a knowledge of the groundwater flow system and how it responds to remedial actions

  7. Recycling endosomes in human cytotoxic T lymphocytes constitute an auxiliary intracellular trafficking pathway for newly synthesized perforin (United States)

    Lesteberg, Kelsey E.; Orange, Jordan S.; Makedonas, George


    Background Although cytotoxic T lymphocytes (CTLs) store perforin within cytoplasmic secretory granules for immediate use, perforin is synthesized anew within hours of TCR stimulation. Previously, we observed new perforin protein at an immunologic synapse independent of secretory lysosomes; herein we aimed to determine how new perforin transits to the synapse if not via lytic granules. Results We analyzed antigen-specific human CTLs via imaging flow cytometry and high-resolution confocal microscopy, with attention to intracellular trafficking components and new perforin. The recycling endosome compartments identified by rab8, rab11a, rab4, and rab37 co-localized with new perforin, as well as the SNAREs vti1b and VAMP4. After ablating the function of the recycling endosome pathway, we observed a relative accumulation of new perforin in rab8 vesicles. Conclusions The recycling endosome pathway may serve as an auxiliary intracellular route for the delivery of new perforin to an immunologic synapse in order to perpetuate a cytotoxic response. PMID:28822075

  8. Structural Basis for the Interaction of the Golgi-Associated Retrograde Protein (GARP) Complex with the t-SNARE Syntaxin 6 (United States)

    Abascal-Palacios, Guillermo; Schindler, Christina; Rojas, Adriana L; Bonifacino, Juan S.; Hierro, Aitor


    Summary The Golgi-Associated Retrograde Protein (GARP) is a tethering complex involved in the fusion of endosome-derived transport vesicles to the trans-Golgi network through interaction with components of the Syntaxin 6/Syntaxin 16/Vti1a/VAMP4 SNARE complex. The mechanisms by which GARP and other tethering factors engage the SNARE fusion machinery are poorly understood. Herein we report the structural basis for the interaction of the human Ang2 subunit of GARP with Syntaxin 6 and the closely related Syntaxin 10. The crystal structure of Syntaxin 6 Habc domain in complex with a peptide from the N terminus of Ang2 shows a novel binding mode in which a di-tyrosine motif of Ang2 interacts with a highly conserved groove in Syntaxin 6. Structure-based mutational analyses validate the crystal structure and support the phylogenetic conservation of this interaction. The same binding determinants are found in other tethering proteins and syntaxins, suggesting a general interaction mechanism. PMID:23932592

  9. Two populations of double minute chromosomes harbor distinct amplicons, the MYC locus at 8q24.2 and a 0.43-Mb region at 14q24.1, in the SW613-S human carcinoma cell line. (United States)

    Guillaud-Bataille, M; Brison, O; Danglot, G; Lavialle, C; Raynal, B; Lazar, V; Dessen, P; Bernheim, A


    High-level amplifications observed in tumor cells are usually indicative of genes involved in oncogenesis. We report here a high resolution characterization of a new amplified region in the SW613-S carcinoma cell line. This cell line contains tumorigenic cells displaying high-level MYC amplification in the form of double minutes (DM(+) cells) and non tumorigenic cells exhibiting low-level MYC amplification in the form of homogeneously staining regions (DM(-) cells). Both cell types were studied at genomic and functional levels. The DM(+) cells display a second amplification, corresponding to the 14q24.1 region, in a distinct population of DMs. The 0.43-Mb amplified and overexpressed region contains the PLEK2, PIGH, ARG2, VTI1B, RDH11, and ZFYVE26 genes. Both amplicons were stably maintained upon in vitro and in vivo propagation. However, the 14q24.1 amplicon was not found in cells with high-level MYC amplification in the form of HSRs, either obtained after spontaneous integration of endogenous DM MYC copies or after transfection of DM(-) cells with a MYC gene expression vector. These HSR-bearing cells are highly tumorigenic. The 14q24.1 amplification may not play a role in malignancy per se but might contribute to maintaining the amplification in the form of DMs. Copyright 2009 S. Karger AG, Basel.

  10. Synaptotagmin 11 interacts with components of the RNA-induced silencing complex RISC in clonal pancreatic β-cells. (United States)

    Milochau, Alexandra; Lagrée, Valérie; Benassy, Marie-Noëlle; Chaignepain, Stéphane; Papin, Julien; Garcia-Arcos, Itsaso; Lajoix, Anne; Monterrat, Carole; Coudert, Laetitia; Schmitter, Jean-Marie; Ochoa, Begoña; Lang, Jochen


    Synaptotagmins are two C2 domain-containing transmembrane proteins. The function of calcium-sensitive members in the regulation of post-Golgi traffic has been well established whereas little is known about the calcium-insensitive isoforms constituting half of the protein family. Novel binding partners of synaptotagmin 11 were identified in β-cells. A number of them had been assigned previously to ER/Golgi derived-vesicles or linked to RNA synthesis, translation and processing. Whereas the C2A domain interacted with the Q-SNARE Vti1a, the C2B domain of syt11 interacted with the SND1, Ago2 and FMRP, components of the RNA-induced silencing complex (RISC). Binding to SND was direct via its N-terminal tandem repeats. Our data indicate that syt11 may provide a link between gene regulation by microRNAs and membrane traffic. Copyright © 2014 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

  11. Soluble N-ethylmaleimide-sensitive factor attachment protein receptors required during Trypanosoma cruzi parasitophorous vacuole development. (United States)

    Cueto, Juan Agustín; Vanrell, María Cristina; Salassa, Betiana Nebaí; Nola, Sébastien; Galli, Thierry; Colombo, María Isabel; Romano, Patricia Silvia


    Trypanosoma cruzi, the etiologic agent of Chagas disease, is an obligate intracellular parasite that exploits different host vesicular pathways to invade the target cells. Vesicular and target soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs) are key proteins of the intracellular membrane fusion machinery. During the early times of T. cruzi infection, several vesicles are attracted to the parasite contact sites in the plasma membrane. Fusion of these vesicles promotes the formation of the parasitic vacuole and parasite entry. In this work, we study the requirement and the nature of SNAREs involved in the fusion events that take place during T. cruzi infection. Our results show that inhibition of N-ethylmaleimide-sensitive factor protein, a protein required for SNARE complex disassembly, impairs T. cruzi infection. Both TI-VAMP/VAMP7 and cellubrevin/VAMP3, two v-SNAREs of the endocytic and exocytic pathways, are specifically recruited to the parasitophorous vacuole membrane in a synchronized manner but, although VAMP3 is acquired earlier than VAMP7, impairment of VAMP3 by tetanus neurotoxin fails to reduce T. cruzi infection. In contrast, reduction of VAMP7 activity by expression of VAMP7's longin domain, depletion by small interfering RNA or knockout, significantly decreases T. cruzi infection susceptibility as a result of a minor acquisition of lysosomal components to the parasitic vacuole. In addition, overexpression of the VAMP7 partner Vti1b increases the infection, whereas expression of a KIF5 kinesin mutant reduces VAMP7 recruitment to vacuole and, concomitantly, T. cruzi infection. Altogether, these data support a key role of TI-VAMP/VAMP7 in the fusion events that culminate in the T. cruzi parasitophorous vacuole development. © 2016 John Wiley & Sons Ltd.

  12. Hydrologic evaluation methodology for estimating water movement through the unsaturated zone at commercial low-level radioactive waste disposal site (United States)

    Meyer, P.D.; Rockhold, M.L.; Nichols, W.E.; Gee, G.W.


    . Several computer simulation codes that were used in the examples (Chapter 6) are discussed with respect to this checklist. The codes used include HELP, UNSAT-H, and VAM3DCG.To provide a defensible estimate of water flow in a LLW disposal facility, the uncertainty associated with model predictions must be considered. Uncertainty arises because of the highly heterogeneous nature of most subsurface environments and the long time frame required in the analysis. Sources of uncertainty in hydrologic evaluation of the unsaturated zone and several approaches for analysis are discussed in Chapter 5. The methods of analysis discussed include a bounding approach, sensitivity analysis, and Monte Carlo simulation.To illustrate the application of the discussion in Chapters 2 through 5, two examples are presented in Chapter 6. The first example is of a below ground vault located in a humid environment. The second example looks at a shallow land burial facility located in an arid environment. The examples utilize actual site-specific data and realistic facility designs. The two examples illustrate the issues unique to humid and arid sites as well as the issues common to all LLW sites. Strategies for addressing the analytical difficulties arising in any complex hydrologic evaluation of the unsaturated zone are demonstrated.The report concludes with some final observations and recommendations.