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Dissertation
Characterization of the YLR190w gene product in the yeast Saccharomyces cerevisiae
Doctor of Philosophy (Ph.D.), Drexel University
Sep 2003
DOI:
https://doi.org/10.17918/00007803
Abstract
Pho85p, a nonessential Cyclin Dependent Kinase (CDK), associates with two cyclins Pc16p and Pc17p, but the function of either complex is unknown. A yeast genome wide two-hybrid screen performed by Fields and co-workers suggested that an uncharacterized open reading frame (ORF) named YLR190w interacts with Pc16p. Recently, YLR190w has been renamed MMR1 at the Saccharomyces Genome Database, therefore, we will refer to YLR190w as MMR1. We have performed a two-hybrid screen to isolate proteins that interact with Mmr1p and have shown that this protein interacts with the carboxyl terminal tail of Myo2p, an essential Class V myosin, and confirmed that Pc16p does indeed interact with Mmr1p. Using epitope tagged versions of Mmr1p we have found that this protein localizes to the area of the bud in yeast. Further studies with synchronous cell cultures show that this localization moves from the point of a newly formed bud through the bud neck. Myo2p has been implicated in playing a key role in the transport of vesicles from the mother cell to the bud and numerous published studies have shown that Myo2p also localizes to sites of polarized cell growth. Our immunofluorescence data show these two proteins co-localize to the same areas. We also examined the protein expression of Mmr1p as it has been shown to localize in a polarized manner and interact with the Pc16p-Pho85p kinase complex. In wild type cells, we found that Mmr1p appears as a single band in early time points after release from [alpha] factor treatment, and at later time points, Mmr1p appears as a doublet. This doublet was subsequently to be shown to be a result of phosphorylation. Results from western analysis of synchronized cells from both pho85[delta] and pc16[delta]pc17[delta] cells suggest that Pho85p is not the kinase responsible for the stage specific phosphorylation of Mmr1p. These results also indicate that Pc16p and Pc17p play a role in this phosphorylation. The work presented here begins to elucidate the function of Mmr1p in the cell.
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Details
- Title
- Characterization of the YLR190w gene product in the yeast Saccharomyces cerevisiae
- Creators
- Kimberly A. Metera
- Contributors
- Lawrence W. Bergman (Advisor) - Drexel University, Drexel University (1970-)
- Awarding Institution
- Drexel University
- Degree Awarded
- Doctor of Philosophy (Ph.D.)
- Publisher
- Drexel University; Philadelphia, Pennsylvania
- Number of pages
- ix, 114 pages
- Resource Type
- Dissertation
- Language
- English
- Academic Unit
- College of Medicine; Drexel University
- Other Identifier
- 991021888990004721