The ATPase motif in RAD51D is required for resistance to DNA interstrand crosslinking agents and interaction with RAD51C

Aaron M. Gruver, Kristi A. Miller, Changanamkandath Rajesh, Phillip G. Smiraldo, Saravanan Kaliyaperumal, Rachel Balder, Katie M. Stiles, Joanna S. Albala, Douglas L. Pittman

Research output: Contribution to journalArticle

21 Scopus citations

Abstract

Homologous recombination (HR) is a mechanism for repairing DNA interstrand crosslinks and double-strand breaks. In mammals, HR requires the activities of the RAD51 family (RAD51, RAD51B, RAD51C, RAD51D, XRCC2, XRCC3 and DMC1), each of which contains conserved ATP binding sequences (Walker Motifs A and B). RAD51D is a DNA-stimulated ATPase that interacts directly with RAD51C and XRCC2. To test the hypothesis that ATP binding and hydrolysis by RAD51D are required for the repair of interstrand crosslinks, site-directed mutations in Walker Motif A were generated, and complementation studies were performed in Rad51d-deficient mouse embryonic fibroblasts. The K113R and K113A mutants demonstrated a respective 96 and 83% decrease in repair capacity relative to wild-type. Further examination of these mutants, by yeast two-hybrid analyses, revealed an 8-fold reduction in the ability to associate with RAD51C whereas interaction with XRCC2 was retained at a level similar to the S111T control. These cell-based studies are the first evidence that ATP binding and hydrolysis by RAD51D are required for efficient HR repair of DNA interstrand crosslinks.

Original languageEnglish (US)
Pages (from-to)433-440
Number of pages8
JournalMutagenesis
Volume20
Issue number6
DOIs
StatePublished - Nov 2005

ASJC Scopus subject areas

  • Genetics
  • Toxicology
  • Genetics(clinical)
  • Health, Toxicology and Mutagenesis

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    Gruver, A. M., Miller, K. A., Rajesh, C., Smiraldo, P. G., Kaliyaperumal, S., Balder, R., Stiles, K. M., Albala, J. S., & Pittman, D. L. (2005). The ATPase motif in RAD51D is required for resistance to DNA interstrand crosslinking agents and interaction with RAD51C. Mutagenesis, 20(6), 433-440. https://doi.org/10.1093/mutage/gei059