Hypersensitivity to mutation and sister-chromatid-exchange induction in CHO cell mutants defective in incising DNA containing UV lesions

Thompson, Larry H.; Brookman, Kerry W.; Dillehay, Larry E.; Mooney, Carolyn L.; Carrano, A.V.

doi:10.1007/bf01543017

Cited by 124 publications

(56 citation statements)

References 30 publications

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“…If not repaired, SSBs pose a threat to both genetic stability and cell survival, resulting in an increased frequency of mutations and chromosome aberrations (15,23,38,50,53,56). The threat posed by unrepaired SSBs most likely reflects their ability to become double-strand breaks during chromosome replication (27).…”

mentioning

confidence: 99%

Central Role for the XRCC1 BRCT I Domain in Mammalian DNA Single-Strand Break Repair

Taylor

Thistlethwaite

Caldecott

2002

Molecular and Cellular Biology

160

127

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The DNA single-strand break repair (SSBR) protein XRCC1 is required for genetic stability and for embryonic viability. XRCC1 possesses two BRCA1 carboxyl-terminal (BRCT) protein interaction domains, denoted BRCT I and II. BRCT II is required for SSBR during G 1 but is dispensable for this process during S/G 2 and consequently for cell survival following DNA alkylation. Little is known about BRCT I, but this domain has attracted considerable interest because it is the site of a genetic polymorphism that epidemiological studies have associated with altered cancer risk. We report that the BRCT I domain comprises the evolutionarily conserved core of XRCC1 and that this domain is required for efficient SSBR during both G 1 and S/G 2 cell cycle phases and for cell survival following treatment with methyl methanesulfonate. However, the naturally occurring human polymorphism in BRCT I supported XRCC1-dependent SSBR and cell survival after DNA alkylation equally well. We conclude that while the BRCT I domain is critical for XRCC1 to maintain genetic integrity and cell survival, the polymorphism does not impact significantly on this function and therefore is unlikely to impact significantly on susceptibility to cancer.

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mentioning

confidence: 99%

Central Role for the XRCC1 BRCT I Domain in Mammalian DNA Single-Strand Break Repair

Taylor

Thistlethwaite

Caldecott

2002

Molecular and Cellular Biology

160

127

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“…More recently it has been shown that hamster cells preferentially repair certain vital sequences that may facilitate UV resistance despite low overall levels of excision repair (3). It has been suggested that Chinese hamster ovary (CHO) cells with mutations affecting the proficiency of DNA repair pathways may be useful for selecting and assessing the expression of homologous and heterologous DNA repair genes (7,8). Such a mutant CHO cell line has been used recently to select and clone putative DNA repair genes from hamster (9) and human (10,11).…”

mentioning

confidence: 99%

Genetic complementation of UV-induced DNA repair in Chinese hamster ovary cells by the denV gene of phage T4.

Valerie¹,

Riel²,

Henderson³

1985

Proc. Natl. Acad. Sci. U.S.A.

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The denV gene of phage T4, encoding the pyrimidine dimer-specific DNA repair enzyme endonuclease V, has been introduced by DNA transfection into the UV-sensitive DNA repair-deficient Chinese hamster ovary (CHO) cell line UV5. Transformants were first selected for resistance to the antibiotic G418 conferred by the neo gene from TnS carried by the same plasmid. A majority of the isolated G418-resistant UV5 clones also showed an increased resistance to 254-nm UV light. One clone, designated I-Al, was found to have an intermediate level of colony-forming ability after UV irradiation when compared to UV5 and wild-type AA8 cells. A Southern blot showed that I-Al carries a single integrated intact copy of the denV gene. Alkaline sucrose gradients revealed a dose-dependent appearance of breaks in the DNA of I-Al cells following UV-irradiation, while unirradiated cells did not exhibit any significant breaks. Analysis of DNA repair by isopycnic sedimentation showed that DNA excision repair by I-Al was at least equal to the level of repair in AA8 cells. These results show that the prokaryotic denV gene can restore UV repair capabilities in vivo to CHO UV5 cells defective in repair of UV-induced damage.

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“…The UVsensitive cell line UV135 was derived from parental CHO AA8 cells as previously described (6). UV135 is deficient in the damage site incision step of ER (60). UV-resistant cell line 38.4.4 was derived by two serial genomic DNA transfers into UV135 cells (41).…”

Section: Methodsmentioning

confidence: 99%

Human ERCC5 cDNA-cosmid complementation for excision repair and bipartite amino acid domains conserved with RAD proteins of Saccharomyces cerevisiae and Schizosaccharomyces pombe.

et al. 1993

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Several human genes related to DNA excision repair (ER) have been isolated via ER cross-species complementation (ERCC) of UV-sensitive CHO cells. We have now isolated and characterized cDNAs for the human ERCC5 gene that complement CHO UV135 cells. The ERCC5 mRNA size is about 4.6 kb. Our available cDNA clones are partial length, and no single clone was active for UV135 complementation. When cDNAs were mixed pairwise with a cosmid clone containing an overlapping 5'-end segment of the ERCC5 gene, DNA transfer produced UV-resistant colonies with 60 to 95% correction of UV resistance relative to either a genomic ERCCS DNA transformant or the CHO AA8 progenitor cells. cDNA-cosmid transformants regained intermediate levels (20 to 45%) of ER-dependent reactivation of a UV-damaged pSVCATgpt reporter plasmid. Our evidence strongly implicates an in situ recombination mechanism in cDNA-cosmid complementation for ER. The complete deduced amino acid sequence of ERCCS was reconstructed from several cDNA clones encoding a predicted protein of 1,186 amino acids. The ERCC5 protein has extensive sequence similarities, in bipartite domains A and B, to products of R4D repair genes of two yeasts, Saccharomyces cerevisiae RAD2 and Schizosaccharomyces pombe rad13. Sequence, structural, and functional data taken together indicate that ERCC5 and its relatives are probable functional homologs. A second locus represented by S. cerevisiae YKL51O and S. pombe rad2 genes is structuraly distinct from the ERCC5 locus but retains vestigial A and B domain similarities. Our analyses suggest that ERCC5 is a nuclear-localized protein with one or more highly conserved helix-loop-helix segments within domains A and B.

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Hypersensitivity to mutation and sister-chromatid-exchange induction in CHO cell mutants defective in incising DNA containing UV lesions

Cited by 124 publications

References 30 publications

Central Role for the XRCC1 BRCT I Domain in Mammalian DNA Single-Strand Break Repair

Central Role for the XRCC1 BRCT I Domain in Mammalian DNA Single-Strand Break Repair

Genetic complementation of UV-induced DNA repair in Chinese hamster ovary cells by the denV gene of phage T4.

Human ERCC5 cDNA-cosmid complementation for excision repair and bipartite amino acid domains conserved with RAD proteins of Saccharomyces cerevisiae and Schizosaccharomyces pombe.

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