Validating the role of Pcl6 in the cell cycle in yeast (Saccharomyces cerevisiae)

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The cell cycle consists of a set of processes that result in the duplication of cellular constituents. Cyclin dependent kinases (Cdks) ensure successful progression of cells through the cell cycle. Cdk activation requires cyclin binding and phosphorylation of its appropriate substrate to tightly regulate cell cycle progression. There are six known Cdks in yeast (Saccharomyces cerevisiae); Cdc28, Kin28, Srb10, Ctk1, Bur1 and Pho85. Cdc28 is the only essential Cdk due to its crucial role in cell cycle progression. The remaining Cdks contribute to cell cycle progression through regulation of gene expression and cellular metabolism. The cyclin partners for Pho85 includes Pcl6. Knowledge about Pcl6 is limited. The goal of this project is explore and validate the role of Pcl6 in yeast cell biology. Based on literature review, we hypothesized that Pcl6 is regulated by components of the Nucleotide Excision Repair (NER) complex in yeast. The NER pathway in yeast is known to repair endogenous DNA damage in yeast cells. Our results confirm our hypothesis. Components of the NER pathway regulate the stability of Pcl6 in our NER mutant strains. Our studies also explore the sensitivity of exogenous DNA damage on the viability of the NER mutants. Our results show cell sensitivity to exogenous DNA damage by 4-Nitroquinoline 1-Oxide, Hydroxyurea, Hydrogen Peroxide, Nocodazole and Methyl-Methane Sulfonate. Future studies will determine the effects of endogenous and exogenous DNA damage on Pcl6 protein level.

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