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PMID: 16717288 Published · epublish English Journal Article Research Support, N.I.H., Extramural Research Support, U.S. Gov't, Non-P.H.S.

Disparate requirements for the Walker A and B ATPase motifs of human RAD51D in homologous recombination.

Nucleic acids research ·Vol. 34 ·No. 9 ·2006-00-00 ·Pages 2833-43

Wiese C, Hinz JM, Tebbs RS, Nham PB, Urbin SS, Collins DW, Thompson LH, Schild D

Abstract

In vertebrates, homologous recombinational repair (HRR) requires RAD51 and five RAD51 paralogs (XRCC2, XRCC3, RAD51B, RAD51C and RAD51D) that all contain conserved Walker A and B ATPase motifs. In human RAD51D we examined the requirement for these motifs in interactions with XRCC2 and RAD51C, and for survival of cells in response to DNA interstrand crosslinks (ICLs). Ectopic expression of wild-type human RAD51D or mutants having a non-functional A or B motif was used to test for complementation of a rad51d knockout hamster CHO cell line. Although A-motif mutants complement very efficiently, B-motif mutants do not. Consistent with these results, experiments using the yeast two- and three-hybrid systems show that the interactions between RAD51D and its XRCC2 and RAD51C partners also require a functional RAD51D B motif, but not motif A. Similarly, hamster Xrcc2 is unable to bind to the non-complementing human RAD51D B-motif mutants in co-immunoprecipitation assays. We conclude that a functional Walker B motif, but not A motif, is necessary for RAD51D's interactions with other paralogs and for efficient HRR. We present a model in which ATPase sites are formed in a bipartite manner between RAD51D and other RAD51 paralogs.

MeSH Terms
Adenosine Triphosphatases/chemistry Amino Acid Motifs Amino Acid Sequence Animals CHO Cells Cricetinae Cricetulus DNA Damage DNA Repair DNA-Binding Proteins/chemistry,genetics,metabolism Genetic Complementation Test Humans Immunoprecipitation Molecular Sequence Data Mutation Rad51 Recombinase/metabolism Recombination, Genetic Two-Hybrid System Techniques
Chemicals
DNA-Binding Proteins RAD51D protein, human Rad51 Recombinase Adenosine Triphosphatases
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Wiese Claudia
Life Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA. cwiese@lbl.gov
Hinz John M
Tebbs Robert S
Nham Peter B
Urbin Salustra S
Collins David W
Thompson Larry H
Schild David
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
1362-4962
DOI
10.1093/nar/gkl366
Email
cwiese@lbl.gov
Published
2006-00-00
Epub
2006-00-22
Pages
2833-43
Language
English
Region
England
NLM ID
0411011
PMCID
PMC1464408
Subset
IM
Grants
NCI NIH HHS · CA112566 · United States
NCI NIH HHS · R01 CA112566 · United States
NCI NIH HHS · CA89405 · United States
NCI NIH HHS · P01 CA092584 · United States
NCI NIH HHS · P01 CA92584 · United States
NCI NIH HHS · R01 CA089405 · United States
External Links
PubMed source DOI: 10.1093/nar/gkl366 PMC Full Text View journal details
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