DNA replication is performed by a group of enzymes called DNA polymerases. A special class of DNA polymerases, called the Y-family polymerases is able to traverse DNA lesions. Cisplatin is a chemotherapeutic agent that damages DNA and has been used to treat cancer. However, many patients become resistant to cisplatin. The potential role of Y-family DNA polymerases in the pathway of chemotherapeutic resistance offers a novel target for inhibition. Growth, purification and expression of the Y-family DNA polymerase, Pol Eta was accomplished by recombinant methods and FPLC (Fast Protein Liquid Chromatography). SDS polyacrylamide gel electrophoresis was used to visualize the success of the purification, and Bradford assay was conducted to quantify the amount of protein purified. A strand-displacement assay as well as primer-template assay was used to verify the protein's ability to synthesize synthetic DNA. Currently, the strand-displacement assay is being used to screen and quantify the roles of potential inhibitors. This work will provide insight into the molecular mechanism of lesion bypass and serve as a platform for potential drug targets against Polh.

Investigation of potential drug inhibitors against DNA polymerase Eta

255th National Meeting (2018)

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