RecO Couples the Assembly of RecA-Mediated DNA Repair Complexes to Replication Forks In Bacillus subtilis
Year: 2011
Type: dissertation
Abstract: All organisms have evolved efficient DNA repair pathways to cope with constant endogenous and exogenous stresses that create nicks, gaps and other lesions in their DNA.Double strand breaks (DSBs) are one of the most lethal forms of DNA damage and are repaired by a form of homologous recombination known as double strand break repair.RecA is a recombinase that is required for homologous recombination and is a positive regulator of the DNA damage response.In addition to being highly conserved amongst bacterial systems, Rad51 and Dcm1 are RecA homologs that are critical for genome maintenance in eukaryotic systems.The mechanism responsible for RecA loading during double strand break repair is well understood in the Gram-negative bacterium Escherichia coli.However, the pathway responsible for RecA loading in B.subtilis is unclear and the recombination mediator proteins involved are not well understood.In this work, we used a RecA-GFP fusion to monitor the localization of RecA-GFP into foci in response to DNA damage in actively replicating cells.Using this approach as an in vivo assay for RecA loading, we found that the protein RecO is obligatory for recruitment of RecA-GFP into foci in replicating cells.Furthermore, we tested several mutant forms of RecO and quantified their effect on RecA-GFP localization in vivo.Taken together, our results show that RecO is required for RecA-GFP localization at the replication fork, suggesting
Author: Eileen R. Brandes
Language: English
Topic: DNA Repair Mechanisms
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: green
OpenAlex ID: W618469233