{"database":"biostudies-literature","file_versions":[],"scores":{"citationCount":0,"reanalysisCount":0,"viewCount":57,"searchCount":0},"additional":{"submitter":["Cockram CA"],"funding":["European Research Council","Medical Research Council","Marie Curie"],"pagination":["E4735-42"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC4553759"],"repository":["biostudies-literature"],"omics_type":["Unknown"],"volume":["112(34)"],"pubmed_abstract":["Understanding molecular mechanisms in the context of living cells requires the development of new methods of in vivo biochemical analysis to complement established in vitro biochemistry. A critically important molecular mechanism is genetic recombination, required for the beneficial reassortment of genetic information and for DNA double-strand break repair (DSBR). Central to recombination is the RecA (Rad51) protein that assembles into a spiral filament on DNA and mediates genetic exchange. Here we have developed a method that combines chromatin immunoprecipitation with next-generation sequencing (ChIP-Seq) and mathematical modeling to quantify RecA protein binding during the active repair of a single DSB in the chromosome of Escherichia coli. We have used quantitative genomic analysis to infer the key in vivo molecular parameters governing RecA loading by the helicase/nuclease RecBCD at recombination hot-spots, known as Chi. Our genomic analysis has also revealed that DSBR at the lacZ locus causes a second RecBCD-mediated DSBR event to occur in the terminus region of the chromosome, over 1 Mb away."],"journal":["Proceedings of the National Academy of Sciences of the United States of America"],"pubmed_title":["Quantitative genomic analysis of RecA protein binding during DNA double-strand break repair reveals RecBCD action in vivo."],"pmcid":["PMC4553759"],"funding_grant_id":["MR/M019160/1","RULE-320823","320823","G0901622","PIOF-GA-2009-254082 - DRIBAC","Studentship and Centenary Award"],"pubmed_authors":["El Karoui M","Cockram CA","Filatenkova M","Leach DR","Danos V"],"view_count":["57"],"additional_accession":[]},"is_claimable":false,"name":"Quantitative genomic analysis of RecA protein binding during DNA double-strand break repair reveals RecBCD action in vivo.","description":"Understanding molecular mechanisms in the context of living cells requires the development of new methods of in vivo biochemical analysis to complement established in vitro biochemistry. A critically important molecular mechanism is genetic recombination, required for the beneficial reassortment of genetic information and for DNA double-strand break repair (DSBR). Central to recombination is the RecA (Rad51) protein that assembles into a spiral filament on DNA and mediates genetic exchange. Here we have developed a method that combines chromatin immunoprecipitation with next-generation sequencing (ChIP-Seq) and mathematical modeling to quantify RecA protein binding during the active repair of a single DSB in the chromosome of Escherichia coli. We have used quantitative genomic analysis to infer the key in vivo molecular parameters governing RecA loading by the helicase/nuclease RecBCD at recombination hot-spots, known as Chi. Our genomic analysis has also revealed that DSBR at the lacZ locus causes a second RecBCD-mediated DSBR event to occur in the terminus region of the chromosome, over 1 Mb away.","dates":{"release":"2015-01-01T00:00:00Z","publication":"2015 Aug","modification":"2024-11-20T01:09:09.73Z","creation":"2019-03-27T01:57:31Z"},"accession":"S-EPMC4553759","cross_references":{"pubmed":["26261330"],"doi":["10.1073/pnas.1424269112"]}}