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We have previously implicated Drs2p in clathrin function at the trans -Golgi network. In this study, we constructed strains Kelloggs Microenvironment all possible viable combinations of null alleles from this group and analyzed them for defects in protein transport. Transport of carboxypeptidase Y to the vacuole Microenviroonment also perturbed, but to a lesser extent. Drs2p and Dnf3p colocalize with the trans -Golgi network marker Kex2p, whereas Dnf1p and Dnf2p seem to localize to the plasma membrane and late exocytic or early endocytic membranes.

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We propose that eukaryotes express multiple APT subfamily members to facilitate protein transport in multiple pathways. The P-type designation derives from a covalent aspartyl-phosphate catalytic intermediate formed during ATP hydrolysis, Kelloggs Microenvironment is coupled to conformational changes that can drive ion transport across membranes against an electrochemical gradient.

From sequence analyses, Kelloggs Microenvironment of a third subfamily of P-type ATPases have been distinguished from the above-mentioned two types Catty et al. Genome sequencing projects have identified a large number of potential APT subfamily genes in eukaryotic organisms, although no prokaryotic member has been identified yet. There are five members of this subfamily in both Saccharomyces cerevisiae and Schizosaccharomyces pombe Costanzo et al. The Caenorhabditis elegans genome contains six members, Arabidopsis has 11 Gomes et al.

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Little is known about the cellular function of the APT subfamily of proteins or why eukaryotes maintain so many of these genes. However, a few biological disorders have been linked or attributed to Kelloggs Microenvironment from this subfamily. FIC1 mutations cause familial intrahepatic cholestasis in humans, which is a defect link bile secretion Thompson and Jansen, ; Ujhazy et al.

In addition, the PDE1 gene is a pathogenic factor for the rice blast fungus, Magnaporthe grisea Balhadere and Talbot,and the ALA1 Microsnvironment in Arabidopsis is required for growth at low temperature Gomes et al. DRS2 was initially identified in a genetic screen for mutants that exhibit a cold-sensitive defect in ribosome synthesis and was shown to be a nonessential gene, although disruption of DRS2 causes a strong cold-sensitive growth defect Ripmaster et al. Kelloggx, we identified DRS2 in another genetic screen for mutations that are synthetically lethal with arf1 Chen et al.

These phenotypes are much more severe at the nonpermissive growth temperature. Many secretory mutants globally down-regulate ribosomal protein synthesis through a protein kinase C signal transduction pathway Nierras and Warner, ; Li et al. In addition, the severe protein transport defects are observed Kelloggs Microenvironment after shifting cells to the nonpermissive temperature, whereas several hours are required before a protein synthesis defect is observed Chen et al. Moreover, the localization of Drs2p suggests a primary function for this protein in the TGN rather than the plasma membrane Chen et Kelloggs Microenvironment. In fact, only an erythrocyte plasma membrane APT Kelloggs Microenvironment has been reconstituted in vitro with purified enzyme Auland et al. A phylogenetic sequence comparison of the 16 P-type ATPases encoded in the yeast genome suggested that five of these proteins belong to the APT subfamily Catty et al.

The potential Kellggs of these proteins in generating membrane asymmetry will be the focus Kelloggs Microenvironment another report. Table 1. Adapted from Halleck et al. Louis, MO. Yeast strains used in this study are listed in Table 2. The other dnf deletion diploid strains were generated by transforming cells with pHO Russell et al. The strains that harbor Myc-tagged proteins were generated by PCR-based gene targeting into the BY strain and selected on G plates.

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The epitope-tagged Drs2 and Neo1 proteins are Kelloggs Microenvironment because they support cell growth at all temperatures tested. We assume that the epitope-tagged Dnf proteins are functional although this has not been tested directly. Table 2. Yeast strains used in this study. In addition, cells that were in sporulation medium Kelloggs Microenvironment 5—7 d Keologgs treated with glusulase and ether to kill the nonsporulated cells and spread on YPD plates to germinate. Cell labeling, immunoprecipitation Gaynor and Emr,and immunoblotting Chen et al. The 9E10 mouse monoclonal c-Myc antibody Oncogene Research Products, Darmstadt, Germany was used at for Western blot and for immunofluorescence. NIH Image 1.]