Quantitative genomic analysis of RecA protein binding during DNA double-strand break repair reveals RecBCD action in vivo
| Autor: | Charlotte A. Cockram, David R. F. Leach, Milana Filatenkova, Vincent Danos, Meriem El Karoui |
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| Přispěvatelé: | University of Edinburgh, School of Informatics [Edimbourg], Analyse Statique par Interprétation Abstraite (ANTIQUE), Département d'informatique - ENS Paris (DI-ENS), École normale supérieure - Paris (ENS-PSL), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de Recherche en Informatique et en Automatique (Inria)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure - Paris (ENS-PSL), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de Recherche en Informatique et en Automatique (Inria)-Centre National de la Recherche Scientifique (CNRS)-Inria Paris-Rocquencourt, Institut National de Recherche en Informatique et en Automatique (Inria), Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche en Informatique et en Automatique (Inria)-École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche en Informatique et en Automatique (Inria)-École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Inria Paris-Rocquencourt, Département d'informatique de l'École normale supérieure (DI-ENS), École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de Recherche en Informatique et en Automatique (Inria)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure - Paris (ENS Paris) |
| Rok vydání: | 2015 |
| Předmět: |
Chromatin Immunoprecipitation
Exodeoxyribonuclease V DNA Repair DNA repair [SDV]Life Sciences [q-bio] RAD51 Genetic recombination 03 medical and health sciences chemistry.chemical_compound Mechanistic modelling [INFO]Computer Science [cs] Homologous recombination 030304 developmental biology Genetics RecBCD 0303 health sciences Multidisciplinary RecA Genome biology 030306 microbiology Helicase Double Strand Break Repair Rec A Recombinases chemistry PNAS Plus biology.protein bacteria DNA DNA Damage |
| Zdroj: | Cockram, C A, Filatenkova, M, Danos, V, El Karoui, M & Leach, D R F 2015, ' Quantitative genomic analysis of RecA protein binding during DNA double-strand break repair reveals RecBCD action in vivo ', Proceedings of the National Academy of Sciences, vol. 112, no. 34, E4735-E4742 . https://doi.org/10.1073/pnas.1424269112 Proceedings of the National Academy of Sciences of the United States of America Proceedings of the National Academy of Sciences of the United States of America, 2015, ⟨10.1073/pnas.1424269112⟩ Proceedings of the National Academy of Sciences of the United States of America, National Academy of Sciences, 2015, ⟨10.1073/pnas.1424269112⟩ |
| ISSN: | 1091-6490 0027-8424 |
| DOI: | 10.1073/pnas.1424269112 |
| Popis: | International audience; 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 recom-bination 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 immunoprecipita-tion 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. homologous recombination | mechanistic modelling | DNA repair | RecA |
| Databáze: | OpenAIRE |
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