Repression of mutagenesis by Rad51D-mediated homologous recombination

John M. Hinz, Robert S. Tebbs, Paul F. Wilson, Peter B. Nham, Edmund P. Salazar, Hatsumi Nagasawa, Salustra S. Urbin, Joel S. Bedford, Larry H. Thompson

Research output: Contribution to journalArticle

61 Citations (Scopus)

Abstract

Homologous recombinational repair (HRR) restores chromatid breaks arising during DNA replication and prevents chromosomal rearrangements that can occur from the misrepair of such breaks. In vertebrates, five Rad51 paralogs are identified that contribute in a nonessential but critical manner to HRR proficiency. We constructed and characterized a knockout of the paralog Rad51D in widely studied CHO cells. The rad51d mutant (clone 51D1) displays sensitivity to a diverse spectrum of induced DNA damage including γ-rays, ultraviolet (UV)-C radiation, and methyl methanesulfonate (MMS), indicating the broad relevance of HRR to genotoxicity. Spontaneous chromatid breaks/gaps and isochromatid breaks are elevated 3- to 12-fold, but the chromosome number distribution remains unchanged. Most importantly, 51D1 cells exhibit a 12-fold-increased rate of hprt mutation, as well as 4- to 10-fold increased rates of gene amplification at the dhfr and CAD loci, respectively. Xrcc3 irs1SF cells from the same parental CHO line show similarly elevated mutagenesis at these three loci. Collectively, these results confirm the a priori expectation that HRR acts in an error-free manner to repress three classes of genetic alterations (chromosomal aberrations, loss of gene function and increased gene expression), all of which are associated with carcinogenesis.

Original languageEnglish (US)
Pages (from-to)1358-1368
Number of pages11
JournalNucleic Acids Research
Volume34
Issue number5
DOIs
StatePublished - 2006
Externally publishedYes

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Recombinational DNA Repair
Homologous Recombination
Mutagenesis
Chromatids
Methyl Methanesulfonate
Chromosomes, Human, Pair 12
CHO Cells
Gene Amplification
Mutation Rate
Ultraviolet Rays
DNA Replication
Chromosome Aberrations
DNA Damage
Vertebrates
Carcinogenesis
Clone Cells
Radiation
Gene Expression
Genes

ASJC Scopus subject areas

  • Genetics

Cite this

Hinz, J. M., Tebbs, R. S., Wilson, P. F., Nham, P. B., Salazar, E. P., Nagasawa, H., ... Thompson, L. H. (2006). Repression of mutagenesis by Rad51D-mediated homologous recombination. Nucleic Acids Research, 34(5), 1358-1368. https://doi.org/10.1093/nar/gkl020

Repression of mutagenesis by Rad51D-mediated homologous recombination. / Hinz, John M.; Tebbs, Robert S.; Wilson, Paul F.; Nham, Peter B.; Salazar, Edmund P.; Nagasawa, Hatsumi; Urbin, Salustra S.; Bedford, Joel S.; Thompson, Larry H.

In: Nucleic Acids Research, Vol. 34, No. 5, 2006, p. 1358-1368.

Research output: Contribution to journalArticle

Hinz, JM, Tebbs, RS, Wilson, PF, Nham, PB, Salazar, EP, Nagasawa, H, Urbin, SS, Bedford, JS & Thompson, LH 2006, 'Repression of mutagenesis by Rad51D-mediated homologous recombination', Nucleic Acids Research, vol. 34, no. 5, pp. 1358-1368. https://doi.org/10.1093/nar/gkl020
Hinz JM, Tebbs RS, Wilson PF, Nham PB, Salazar EP, Nagasawa H et al. Repression of mutagenesis by Rad51D-mediated homologous recombination. Nucleic Acids Research. 2006;34(5):1358-1368. https://doi.org/10.1093/nar/gkl020
Hinz, John M. ; Tebbs, Robert S. ; Wilson, Paul F. ; Nham, Peter B. ; Salazar, Edmund P. ; Nagasawa, Hatsumi ; Urbin, Salustra S. ; Bedford, Joel S. ; Thompson, Larry H. / Repression of mutagenesis by Rad51D-mediated homologous recombination. In: Nucleic Acids Research. 2006 ; Vol. 34, No. 5. pp. 1358-1368.
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