xii, 45 p. : col. ill. / Regulated acidification of intercellular organelles and vesicles is essential for many cellular processes, from basic metabolism and protein sorting to synapse function and developmental signaling. These diverse processes are driven by spatiotemporal regulation of the V-ATPase, the cellular H + -pump. In yeast and higher eukaryotes V-ATPase localization is directed by the 100-kDa "a" subunit, and many human diseases are linked to mutations in "a". Saccharomyces cerevisiae contains two "a" isoforms, VPH1 and STV1, with all other V-ATPase subunits encoded for by single genes. The V-ATPase contains only one "a" subunit per complex. Complexes that contain Vph1p localize to the vacuole (lysosome), and Stv1p-containing complexes localize to the late Golgi/endosome. Here I present a set of STV1 mutants that are disrupted for Golgi retention but not V-ATPase assembly or enzymatic function. Using a forward genetic screen I defined multiple residues within a 39 amino-acid region of Stv1p that are necessary for Stv1p retention to the Golgi. The residues most strongly affecting Golgi localization are present in a small STV1 -specific insertion of eight residues, suggesting they may bind directly to sorting machinery. However, I also find that Stv1p/Vph1p chimeras containing the STV1 -specific insertion are not sufficient to direct Golgi retention in both minimal (13AA) and expanded (49AA) contexts. I conclude that the Stv1p Golgi retention signal is composed of a complex binding surface, of which the central element is a short peptide rich in amino acids with aromatic side chains. / Committee in charge: Bruce Bowerman, Chairperson;
Tom H. Stevens, Advisor;
Karen Guillemin, Member;
George F. Sprague Jr., Member
Kenneth E. Prehoda Outside Member
| Identifer | oai:union.ndltd.org:uoregon.edu/oai:scholarsbank.uoregon.edu:1794/11646 |
| Date | 06 1900 |
| Creators | Cronan, Glen Emerson, 1977- |
| Publisher | University of Oregon |
| Source Sets | University of Oregon |
| Language | en_US |
| Detected Language | English |
| Type | Thesis |
| Relation | University of Oregon theses, Dept. of Biology, Ph. D., 2011; |
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