Periodic Reporting for period 1 - DNA Pol III (How single DNA polymerases make decisions during proofreading)
Período documentado: 2016-04-01 hasta 2018-03-31
This goal of this work is to study how single DNA polymerase enzymes carry out the molecular acrobatic act of proofreading. In order to do this, we have developed a single-molecule system where we can watch single DNA polymerase enzymes perform proofreading in real time. We have monitored the kinetics and protein binding times for DNA polymrease III during the course of a proofreading reaction. We have studied how proofreading changes as a function of the type of error in DNA. Further, we have determined that the conformational dynamics during proofreading for DNA polymerase III change in comparing the wild type and a mutant enzyme that is deficient in proofreading. It is our hope that this work will shift the knowledge frontier by advancing our understanding of proofreading during DNA replication in order to better realise the relationship between DNA mutations and the development of human diseases like cancer.