Rev1 DNA Polymerase Repair Mechanism and Optical Tweezer Measurements on Damaged DNA*

Replication of damaged DNA leads to cancer. The nucleotide arrangement of DNA contributes to its strength, which can be altered by damage. One mechanism by which the body repairs damage is DNA polymerases, specifically Rev1. Rev1 is an enzyme that facilitates the replication or repair of specific damaged or absent nucleotides in DNA and is a component of the body's ability to prevent mutated DNA from replication and cancer. We hypothesize two amino acids in the active site of Rev1 facilitate its unique Y-family polymerase repair mechanisms using protein purification and x-ray crystallography.



In conjunction with understanding Rev1's ability to repair damaged DNA, we are also investigating the inherent structural properties of DNA and how damage alters those properties. A comparison between healthy DNA and unrepaired DNA is crucial. Force measurements necessary to break healthy and unhealthy DNA can be acquired using optical tweezers. The force measurements will provide numerical data for a previously unquantifiable event. Our group built a custom optical tweezer setup for undergraduate institutions lacking sufficient funds to buy optics equipment. These force measurements will help us better comprehend the importance of Rev1's DNA repair mechanism.