Date of Award

12-1-2014

Degree Name

Doctor of Philosophy

Department

Molecular Biology, Microbiology and Biochemistry

First Advisor

Kadyrov, Farid

Abstract

Mismatch repair corrects errors made during DNA replication and inactive mismatch repair is associated with Lynch Syndrome and sporadic cancer. Genome replication in eukaryotes is accompanied by chromatin formation. The first step in chromatin establishment is nucleosome assembly, that starts with histone tetramer deposition. It is not clear how three important cellular processes: genome replication, mismatch repair and nucleosome assembly are coordinated. Here we analyzed human mismatch repair in the presence of histone deposition in a reconstituted system. We showed that mismatch repair factor inhibits nucleosome assembly on the DNA region with the replicative error. Such a mechanism is important, since in this way DNA with errors remains accessible for mismatch repair system to perform the repair. The DNA synthesis step in mismatch repair is performed by DNA polymerase. Eukaryotes possess two major replicative DNA Polymerases: DNA Polymerase delta and DNA Polymerase epsilon. DNA polymerase delta is involved in mismatch repair. However, it was unknown whether DNA polymerase epsilon can also work in mismatch repair. Here we analyzed human mismatch repair with DNA Polymerase delta and DNA Polymerase epsilon in the environment of histone deposition. Our results indicated that repair activity with both polymerases was activated by histone deposition. Here it was first shown that human DNA Polymerase epsilon performs DNA synthesis during mismatch repair in vitro. Importantly, recent studies have revealed association of Polymerase epsilon mutations with cancer. Since our data showed activity of DNA Polymerase epsilon in mismatch repair, a possible tumor development mechanism may involve inactivation of mismatch repair due to Polymerase epsilon mutations. Overall, our study expanded the understanding of the mechanism of human mismatch repair in the chromatin environment.

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