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Mechanistic Understanding Of PCNA-based Interactions: The Interplay Between CAF-I and Rrm3p for the DNA Sliding Clamp PCNA

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Title: Mechanistic Understanding Of PCNA-based Interactions: The Interplay Between CAF-I and Rrm3p for the DNA Sliding Clamp PCNA
Author: Dhavarasa, Piriththiv
Department: Department of Molecular and Cellular Biology
Program: Molecular and Cellular Biology
Advisor: Yankulov, Krassimir
Abstract: The Proliferating Cell Nuclear Antigen (PCNA) is a homotrimeric ring that encircles DNA at the replication fork and acts as a docking hub for numerous replication related proteins. Chromatin Assembly Factor I (CAF-I) and rDNA Recombination Mutation 3 (RRM3) have been implicated in the conversions of the epigenetic state during DNA replication and interact with PCNA through a PCNA-Interacting Peptide (PIP) sequence. The exchange of these factors at the replication fork must play a key role in the coordination of successful replication and the maintenance of chromatin structure. I used a co-immunoprecipitation assay and a GST-based pulldown assay to determine the interaction profile of these proteins and observed that a co-operative tri-partite complex seems plausible. Furthermore, I investigated the significance of CDC7 and CDC28 phosphorylation on CAF-I association with PCNA using a Yeast-Two Hybrid assay. My results show that eliminating putative phosphorylation sites reduced association to differing degrees.
URI: http://hdl.handle.net/10214/12089
Date: 2017-12
Rights: Attribution-NonCommercial-ShareAlike 2.5 Canada


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Attribution-NonCommercial-ShareAlike 2.5 Canada Except where otherwise noted, this item's license is described as Attribution-NonCommercial-ShareAlike 2.5 Canada