Functional characterization of remodeling complex Ino80p

College Of Staten Island, New York NY

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Abstract

DESCRIPTION (provided by applicant): Chromatin structure can play a decisive role in gene regulation by virtue of its dynamic nature upon gene activation. The dynamic changes of chromatin structure have been attributed to ATP-dependent chromatin remodeling complexes. These complexes function by increasing nucleosome mobility and are clearly implicated in transcription. However, the detailed mechanism of gene activation directed by chromatin remodeling complexes is still unclear. In light of the clinical perspective, the newly discovered chromatin remodeling complex Ino80p has been shown to exhibit DNA-dependent ATPase activity as well as DNA damage repair. As such, our long-term objective is to understand the dynamics and mechanisms underlying chromatin remodeling directed by Ino80p and its role in DNA damage repair. In this study, we plan to employ a novel method to isolate native chromatin from yeast cells and use it for monomer extension analysis, chromatin immunoprecipitation analysis, topological analysis and restriction enzyme accessibility analysis to examine the role of Ino80p in INO1 gene induction. Results of the work proposed here will provide a better understanding of the mechanism of gene activation, and the role of Ino80p in gene expression. Most importantly, it will improve our knowledge towards the mechanistic model of chromatin remodeling during gene activation and have implications in DNA damage repair.