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PMID: 11283264 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Chromosome instability and defective recombinational repair in knockout mutants of the five Rad51 paralogs.

Molecular and cellular biology ·Vol. 21 ·No. 8 ·2001-04-00 ·Pages 2858-66

Takata M, Sasaki MS, Tachiiri S, Fukushima T, Sonoda E, Schild D, Thompson LH, Takeda S

Abstract

The Rad51 protein, a eukaryotic homologue of Escherichia coli RecA, plays a central role in both mitotic and meiotic homologous DNA recombination (HR) in Saccharomyces cerevisiae and is essential for the proliferation of vertebrate cells. Five vertebrate genes, RAD51B, -C, and -D and XRCC2 and -3, are implicated in HR on the basis of their sequence similarity to Rad51 (Rad51 paralogs). We generated mutants deficient in each of these proteins in the chicken B-lymphocyte DT40 cell line and report here the comparison of four new mutants and their complemented derivatives with our previously reported rad51b mutant. The Rad51 paralog mutations all impair HR, as measured by targeted integration and sister chromatid exchange. Remarkably, the mutant cell lines all exhibit very similar phenotypes: spontaneous chromosomal aberrations, high sensitivity to killing by cross-linking agents (mitomycin C and cisplatin), mild sensitivity to gamma rays, and significantly attenuated Rad51 focus formation during recombinational repair after exposure to gamma rays. Moreover, all mutants show partial correction of resistance to DNA damage by overexpression of human Rad51. We conclude that the Rad51 paralogs participate in repair as a functional unit that facilitates the action of Rad51 in HR.

MeSH Terms
Animals Avian Proteins Cell Line Cell Survival/drug effects,radiation effects Chickens Chromosomes/genetics Cross-Linking Reagents/pharmacology DNA Repair DNA-Binding Proteins/genetics,metabolism Gamma Rays Gene Deletion Gene Targeting Genetic Complementation Test Humans Phenotype Rad51 Recombinase Recombination, Genetic
Chemicals
Avian Proteins Cross-Linking Reagents DNA-Binding Proteins RAD51 protein, Gallus gallus RAD51 protein, human Rad51 Recombinase
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Takata M
CREST Research Project, Radiation Genetics, Faculty of Medicine, Kyoto University, Sakyo-ku, Kyoto 606-8501, Japan.
Sasaki M S
Tachiiri S
Fukushima T
Sonoda E
Schild D
Thompson L H
Takeda S
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.21.8.2858-2866.2001
Published
2001-04-00
Pages
2858-66
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC86915
Subset
IM
Grants
NIGMS NIH HHS · GM30990 · United States
External Links
PubMed source DOI: 10.1128/MCB.21.8.2858-2866.2001 PMC Full Text View journal details
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