Originally published as Genetics Published Articles Ahead of Print on July 14, 2005.

Genetics, Vol. 171, 873-883, November 2005, Copyright © 2005
doi:10.1534/genetics.105.045906

The RuvAB Branch Migration Translocase and RecU Holliday Junction Resolvase Are Required for Double-Stranded DNA Break Repair in Bacillus subtilis

Humberto Sanchez^*, Dawit Kidane^,1, Patricia Reed, Fiona A. Curtis, M. Castillo Cozar^*, Peter L. Graumann^,1, Gary J. Sharples and Juan C. Alonso^*,2

^* Department of Microbial Biotechnology, Centro Nacional de Biotecnología, Consejo Superior de Investigaciones Científicas, 28049 Madrid, Spain, Biochemie, Fachbereich Chemie, Philipps-Universität Marburg, 35032 Marburg, Germany and Centre for Infectious Diseases, Wolfson Research Institute, University of Durham, Stockton-on-Tees TS17 6BH, United Kingdom

² Corresponding author: Departmento de Biotecnología Microbiana, Centro Nacional de Biotecnología, CSIC, Campus Universidad Autónoma de Madrid, C/Darwin 3, Cantoblanco, 28049 Madrid, Spain.
E-mail: jcalonso{at}cnb.uam.es

In models of Escherichia coli recombination and DNA repair, the RuvABC complex directs the branch migration and resolution of Holliday junction DNA. To probe the validity of the E. coli paradigm, we examined the impact of mutations in ruvAB and recU (a ruvC functional analog) on DNA repair. Under standard transformation conditions we failed to construct ruvAB recG, recU ruvAB, recU recG, or recU recJ strains. However, ruvAB could be combined with addAB (recBCD), recF, recH, recS, recQ, and recJ mutations. The ruvAB and recU mutations rendered cells extremely sensitive to DNA-damaging agents, although less sensitive than a recA strain. When damaged cells were analyzed, we found that RecU was recruited to defined double-stranded DNA breaks (DSBs) and colocalized with RecN. RecU localized to these centers at a later time point during DSB repair, and formation was dependent on RuvAB. In addition, expression of RecU in an E. coli ruvC mutant restored full resistance to UV light only when the ruvAB genes were present. The results demonstrate that, as with E. coli RuvABC, RuvAB targets RecU to recombination intermediates and that all three proteins are required for repair of DSBs arising from lesions in chromosomal DNA.

This article has been cited by other articles:

				P. P. Cardenas, B. Carrasco, H. Sanchez, G. Deikus, D. H Bechhofer, and J. C Alonso Bacillus subtilis polynucleotide phosphorylase 3'-to-5' DNase activity is involved in DNA repair Nucleic Acids Res., May 11, 2009; (2009) gkp314v1. [Abstract] [Full Text] [PDF]

				N. C. Stephanou, F. Gao, P. Bongiorno, S. Ehrt, D. Schnappinger, S. Shuman, and M. S. Glickman Mycobacterial Nonhomologous End Joining Mediates Mutagenic Repair of Chromosomal Double-Strand DNA Breaks J. Bacteriol., July 15, 2007; 189(14): 5237 - 5246. [Abstract] [Full Text] [PDF]

				V. S. Lioy, M. T. Martin, A. G. Camacho, R. Lurz, H. Antelmann, M. Hecker, E. Hitchin, Y. Ridge, J. M. Wells, and J. C. Alonso pSM19035-encoded {zeta} toxin induces stasis followed by death in a subpopulation of cells. Microbiology, August 1, 2006; 152(Pt 8): 2365 - 2379. [Abstract] [Full Text] [PDF]

				H. Sanchez, D. Kidane, M. Castillo Cozar, P. L. Graumann, and J. C. Alonso Recruitment of Bacillus subtilis RecN to DNA Double-Strand Breaks in the Absence of DNA End Processing J. Bacteriol., January 15, 2006; 188(2): 353 - 360. [Abstract] [Full Text] [PDF]

				G. Lopez-Torrejon, M. I. Martinez-Jimenez, and S. Ayora Role of LrpC from Bacillus subtilis in DNA transactions during DNA repair and recombination Nucleic Acids Res., January 10, 2006; 34(1): 120 - 129. [Abstract] [Full Text] [PDF]

International Access Link

Online ISSN: 1943-2361  Print ISSN: 0016-6731
Copyright 2009 by the Genetics Society of America
phone: 412-268-1812   fax: 412-268-1813   email: genetics-gsa{at}andrew.cmu.edu