Interplay between human DNA repair proteins at a unique double-strand break in vivo Journal Article
| Authors: | Rodrigue, A.; Lafrance, M.; Gauthier, M. C.; McDonald, D.; Hendzel, M.; West, S. C.; Jasin, M.; Masson, J. Y. |
| Article Title: | Interplay between human DNA repair proteins at a unique double-strand break in vivo |
| Abstract: | DNA repair by homologous recombination is essential for preserving genomic integrity. The RAD51 paralogs (RAD51B, RAD51C, RAD51D, XRCC2 and XRCC3) play important roles in this process. In this study, we show that human RAD51 interacts with RAD51C-XRCC3 or RAD51B-C-D-XRCC2. In addition to being critical for RAD51 focus formation, RAD51C localizes to DNA damage sites. Inhibition of RAD51C results in a decrease in cellular proliferation consistent with a role in repairing double-strand breaks (DSBs) that occur naturally. To monitor a single DNA repair event, we developed immunofluorescence and chromatin immunoprecipitation (ChIP) methods on human cells where a unique DSB can be created in vivo. Using this system, we observed a single focus of RAD51C, RAD51 and 53BP1, which colocalized with γ-H2AX. ChIPs revealed that endogenous human RAD51, RAD51C, RAD51D, XRCC2, XRCC3 and MRE11 proteins are recruited in the S-G2 phase of the cell cycle, while Ku80 is recruited during G1. We propose that RAD51C ensures a tight regulation of RAD51 assembly during DSB repair and plays a direct role in repairing DSBs in vivo. © 2006 European Molecular Biology Organization | All Rights Reserved. |
| Keywords: | unclassified drug; human cell; dna-binding proteins; protein localization; cell proliferation; mre11 protein; dna damage; homologous recombination; cell survival; cells, cultured; cell cycle; cell cycle s phase; dna repair; embryo; protein assembly; protein protein interaction; protein; in vivo study; immunofluorescence; dna strand breakage; dna; double stranded dna; chromatin immunoprecipitation; recombination, genetic; intracellular signaling peptides and proteins; phosphoproteins; cell cycle g2 phase; histones; cell cycle g1 phase; rad51 protein; genome stability; rad51 recombinase; nucleic acid binding protein; protein 53bp1; protein ku80; rad51; rad51 paralogs; protein rad51b; protein rad51c; protein rad51d; protein xrcc2; protein xrcc3 |
| Journal Title: | EMBO Journal |
| Volume: | 25 |
| Issue: | 1 |
| ISSN: | 0261-4189 |
| Publisher: | Wiley Blackwell |
| Date Published: | 2006-01-11 |
| Start Page: | 222 |
| End Page: | 231 |
| Language: | English |
| DOI: | 10.1038/sj.emboj.7600914 |
| PUBMED: | 16395335 |
| PROVIDER: | scopus |
| PMCID: | PMC1356357 |
| DOI/URL: | |
| Notes: | --- - "Export Date: 4 June 2012" - "CODEN: EMJOD" - "Source: Scopus" |
