Biochemical and Genetic Studies of Genomic Stability

摘要: Genomes face a constant barrage of threats from endogenous and exogenous sources. The need to maintain fidelity while replicating the entire genome during each cell division necessitates dynamic cadre proteins protein complexes that participate in DNA replication process. Furthermore, can be damaged all phases cell’s life damage must recognized repaired way preserves genetic information. These studies focus on one enzyme at nexus repair, helicase/nuclease Dna2. We show Dna2 possesses novel ATP/Mn2+ dependent flap endo/exonuclease activity end-independent endonuclease is inhibited by Replication Protein A. regulation context global response great interest. To end, we explored relationship sensor kinase Mec1. find phosphorylated Mec1 following its N-terminal domain. then extended these yeast higher eukaryotes utilizing Xenopus free extract system. Using simulated double strand breaks (DSBs), constructed timeline processing steps required for homologous recombination mediated repair. This strategy using extracts also place chromatin replication, physically interacting with other involved lagging replication. Taken together, biochemical elucidate multiple roles plays order ensure genomic stability.