ATP hydrolysis by mammalian RAD51 has a key role during homology-directed DNA repair Journal Article

Authors: Stark, J. M.; Hu, P.; Pierce, A. J.; Moynahan, M. E.; Ellis, N.; Jasin, M.
Article Title: ATP hydrolysis by mammalian RAD51 has a key role during homology-directed DNA repair
Abstract: Disruption of the gene encoding RAD51, the protein that catalyzes strand exchange during homologous recombination, leads to the accumulation of chromosome breaks and lethality in vertebrate cells. As RAD51 is implicated in BRCA1- and BRCA2-mediated tumor suppression as well as cellular viability, we have begun a functional analysis of a defined RAD51 mutation in mammalian cells. By using a dominant negative approach, we generated a mouse embryonic stem cell line that expresses an ATP hydrolysis-defective RAD51 protein, hRAD51-K133R, at comparable levels to the endogenous wild-type RAD51 protein, whose expression is retained in these cells. We found that these cells have increased sensitivity to the DNA-damaging agents mitomycin C and ionizing radiation and also exhibit a decreased rate of spontaneous sister-chromatid exchange. By using a reporter for the repair of a single chromosomal double-strand break, we also found that expression of the hRAD51-K133R protein specifically inhibits homology-directed double-strand break repair. Furthermore, expression of a BRC repeat from BRCA2, a peptide inhibitor of an early step necessary for strand exchange, exacerbates the inhibition of homology-directed repair in the hRAD51-K133R expressing cell line. Thus, ATP hydrolysis by RAD51 has a key role in various types of DNA repair in mammalian cells.
Keywords: controlled study; protein expression; human cell; dna-binding proteins; dna recombination; mammalia; animals; mice; dna damage; homologous recombination; dna repair; genes; brca1 protein; brca2 protein; vertebrata; dna; tumors; gene disruption; recombination, genetic; ionizing radiation; mitomycin c; sister chromatid exchange; base sequence; catalysis; dna primers; adenosine triphosphate; biochemistry; hydrolysis; gene encoding; rad51 protein; rad51 recombinase; cell dna; adenosinetriphosphate; human; priority journal; article
Journal Title: Journal of Biological Chemistry
Volume: 277
Issue: 23
ISSN: 0021-9258
Publisher: American Society for Biochemistry and Molecular Biology  
Date Published: 2002-06-07
Start Page: 20185
End Page: 20194
Language: English
DOI: 10.1074/jbc.M112132200
PUBMED: 11923292
PROVIDER: scopus
Notes: Export Date: 14 November 2014 -- Source: Scopus
Citation Impact
MSK Authors
  1. Mary Ellen Moynahan
    105 Moynahan
  2. Peng Hu
    7 Hu
  3. Jeremy M Stark
    8 Stark
  4. Andrew J Pierce
    11 Pierce
  5. Nathan A Ellis
    74 Ellis
  6. Maria Jasin
    231 Jasin