Electronic Thesis and Dissertation Repository

Degree

Master of Science

Program

Biochemistry

Supervisor(s)

Dr. Caroline Schild-Poulter

Abstract

Ku is a key component of the Non-Homologous End Joining DNA repair pathway. Recently, a function for Ku in DNA damage response (DDR) signalling was identified through studies exploring a Ku70 S155D phosphomimetic mutant. We hypothesize that Ku70 S155D mimics phosphorylation of Ku70 in response to DNA damage, and that Ku S155 phosphorylation inhibits Aurora B and causes sustained DDR activation. In this study we show that the S155D mutant is competent for heterodimerization, and its expression does not induce DNA damage. Phosphorylated Ku70 associates with Aurora B by co-immunoprecipitation and this association was demonstrated in situ with Ku70 S155D. Additionally, we demonstrate that the Ku70 S155D vWA domain is sufficient to inhibit Aurora B in an in vitro kinase assay. Finally, Aurora B inhibitor treatment of Ku70 S155D cells does not increase the prevalence of a DDR marker gammaH2AX. This work suggests that Ku70 S155 phosphorylation leads to an inhibition of Aurora B and DDR activation.


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Biochemistry Commons

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