Transformation of normal cells to cancer cells is a result of cumulated errors within specific regulatory genes. How DNA obtains errors in these genes has been an area of cancer research that is under intense examination. This research project is focused on investigating a special pathway, named translesion DNA synthesis (TLS). TLS is a process in which DNA damage is copied by a novel class of enzymes. Rev1 is a member of these enzymes recruited to bypass certain DNA adducts. The purpose of this project is to measure and quantify Rev1’s ability to perform DNA replication across liver carcinogen 1,N-6 etheno deoxyadenosine (etheno-dA). I have expressed and purified Rev1 to homogeneity using affinity chromatography and size exclusion chromatography. A template-primer insertion analysis assay was done to determine the ability of Rev1 to replicate DNA that has been modified with etheno-dA. Primer-template analysis reveals that Rev1 incorporates the incorrect nucleotide dCTP across from the etheno-dA lesion. The research will provide molecular details of DNA lesion bypass and will provide a more complete understanding of carcinogenesis.

Characterization of the lesion bypass activity of Rev1 with the carcinogenic DNA adduct 1,N-6 etheno deoxyadenosine

251st National Meeting (2016)

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