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Mol Cell Biol. 1990 December; 10(12): 6160–6171.
PMCID: PMC362891

Molecular cloning of the human XRCC1 gene, which corrects defective DNA strand break repair and sister chromatid exchange.


We describe the cloning and function of the human XRCC1 gene, which is the first mammalian gene isolated that affects cellular sensitivity to ionizing radiation. The CHO mutant EM9 has 10-fold-higher sensitivity to ethyl methanesulfonate, 1.8-fold-higher sensitivity to ionizing radiation, a reduced capacity to rejoin single-strand DNA breaks, and a 10-fold-elevated level of sister chromatid exchange compared with the CHO parental cells. The complementing human gene was cloned from a cosmid library of a tertiary transformant. Two cosmid clones produced transformants that showed approximately 100% correction of the repair defect in EM9 cells, as determined by the kinetics of strand break repair, cell survival, and the level of sister chromatid exchange. A nearly full-length clone obtained from the pcD2 human cDNA expression library gave approximately 80% correction of EM9, as determined by the level of sister chromatid exchange. Based on an analysis of the nucleotide sequence of the cDNA insert compared with that of the 5' end of the gene from a cosmid clone, the cDNA clone appeared to be missing approximately 100 bp of transcribed sequence, including 26 nucleotides of coding sequence. The cDNA probe detected a single transcript of approximately 2.2 kb in HeLa polyadenylated RNA by Northern (RNA) blot hybridization. From the open reading frame and the positions of likely start sites for transcription and translation, the size of the putative XRCC1 protein is 633 amino acids (69.5 kDa). The size of the XRCC1 gene is 33 kb, as determined by localizing the endpoints on a restriction endonuclease site map of one cosmid clone. The deduced amino acid sequence did not show significant homology with any protein in the protein sequence data bases examined.

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  • Bardwell L. The mutagenic and carcinogenic effects of gene transfer. Mutagenesis. 1989 Jul;4(4):245–253. [PubMed]
  • Birnboim HC, Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. [PMC free article] [PubMed]
  • Birnstiel ML, Busslinger M, Strub K. Transcription termination and 3' processing: the end is in site! Cell. 1985 Jun;41(2):349–359. [PubMed]
  • Boyd JB, Mason JM, Yamamoto AH, Brodberg RK, Banga SS, Sakaguchi K. A genetic and molecular analysis of DNA repair in Drosophila. J Cell Sci Suppl. 1987;6:39–60. [PubMed]
  • Breathnach R, Chambon P. Organization and expression of eucaryotic split genes coding for proteins. Annu Rev Biochem. 1981;50:349–383. [PubMed]
  • Brown WM. Polymorphism in mitochondrial DNA of humans as revealed by restriction endonuclease analysis. Proc Natl Acad Sci U S A. 1980 Jun;77(6):3605–3609. [PubMed]
  • Carrano AV, Minkler JL, Dillehay LE, Thompson LH. Incorporated bromodeoxyuridine enhances the sister-chromatid exchange and chromosomal aberration frequencies in an EMS-sensitive Chinese hamster cell line. Mutat Res. 1986 Sep;162(2):233–239. [PubMed]
  • Chen C, Okayama H. High-efficiency transformation of mammalian cells by plasmid DNA. Mol Cell Biol. 1987 Aug;7(8):2745–2752. [PMC free article] [PubMed]
  • Corsaro CM, Pearson ML. Enhancing the efficiency of DNA-mediated gene transfer in mammalian cells. Somatic Cell Genet. 1981 Sep;7(5):603–616. [PubMed]
  • Dale RM, McClure BA, Houchins JP. A rapid single-stranded cloning strategy for producing a sequential series of overlapping clones for use in DNA sequencing: application to sequencing the corn mitochondrial 18 S rDNA. Plasmid. 1985 Jan;13(1):31–40. [PubMed]
  • Dang CV, Lee WM. Identification of the human c-myc protein nuclear translocation signal. Mol Cell Biol. 1988 Oct;8(10):4048–4054. [PMC free article] [PubMed]
  • Debenham PG, Jones NJ, Webb MB. Vector-mediated DNA double-strand break repair analysis in normal, and radiation-sensitive, Chinese hamster V79 cells. Mutat Res. 1988 May;199(1):1–9. [PubMed]
  • Dillehay LE, Thompson LH, Carrano AV. DNA-strand breaks associated with halogenated pyrimidine incorporation. Mutat Res. 1984 Mar-Apr;131(3-4):129–136. [PubMed]
  • Friedberg EC. Deoxyribonucleic acid repair in the yeast Saccharomyces cerevisiae. Microbiol Rev. 1988 Mar;52(1):70–102. [PMC free article] [PubMed]
  • Fuller LF, Painter RB. A Chinese hamster ovary cell line hypersensitive to ionizing radiation and deficient in repair replication. Mutat Res. 1988 Mar;193(2):109–121. [PubMed]
  • Giaccia A, Weinstein R, Hu J, Stamato TD. Cell cycle-dependent repair of double-strand DNA breaks in a gamma-ray-sensitive Chinese hamster cell. Somat Cell Mol Genet. 1985 Sep;11(5):485–491. [PubMed]
  • Hoy CA, Fuscoe JC, Thompson LH. Recombination and ligation of transfected DNA in CHO mutant EM9, which has high levels of sister chromatid exchange. Mol Cell Biol. 1987 May;7(5):2007–2011. [PMC free article] [PubMed]
  • Hoy CA, Salazar EP, Thompson LH. Rapid detection of DNA-damaging agents using repair-deficient CHO cells. Mutat Res. 1984 Oct;130(5):321–332. [PubMed]
  • Jeggo PA, Kemp LM. X-ray-sensitive mutants of Chinese hamster ovary cell line. Isolation and cross-sensitivity to other DNA-damaging agents. Mutat Res. 1983 Dec;112(6):313–327. [PubMed]
  • Jones NJ, Cox R, Thacker J. Isolation and cross-sensitivity of X-ray-sensitive mutants of V79-4 hamster cells. Mutat Res. 1987 May;183(3):279–286. [PubMed]
  • Jones NJ, Cox R, Thacker J. Six complementation groups for ionising-radiation sensitivity in Chinese hamster cells. Mutat Res. 1988 Mar;193(2):139–144. [PubMed]
  • Jones NJ, Stewart SA, Thompson LH. Biochemical and genetic analysis of the Chinese hamster mutants irs1 and irs2 and their comparison to cultured ataxia telangiectasia cells. Mutagenesis. 1990 Jan;5(1):15–23. [PubMed]
  • Karn J, Matthes HW, Gait MJ, Brenner S. A new selective phage cloning vector, lambda 2001, with sites for XbaI, BamHI, HindIII, EcoRI, SstI and XhoI. Gene. 1984 Dec;32(1-2):217–224. [PubMed]
  • Kemp LM, Sedgwick SG, Jeggo PA. X-ray sensitive mutants of Chinese hamster ovary cells defective in double-strand break rejoining. Mutat Res. 1984 Nov-Dec;132(5-6):189–196. [PubMed]
  • MacInnes MA, Mudgett JS. Cloning of the functional human excision repair gene ERCC-5: potential gene regulatory features conserved with other human repair genes. Prog Clin Biol Res. 1990;340A:265–274. [PubMed]
  • Mansour SL, Thomas KR, Capecchi MR. Disruption of the proto-oncogene int-2 in mouse embryo-derived stem cells: a general strategy for targeting mutations to non-selectable genes. Nature. 1988 Nov 24;336(6197):348–352. [PubMed]
  • Messing J. New M13 vectors for cloning. Methods Enzymol. 1983;101:20–78. [PubMed]
  • Mohrenweiser HW, Carrano AV, Fertitta A, Perry B, Thompson LH, Tucker JD, Weber CA. Refined mapping of the three DNA repair genes, ERCC1, ERCC2, and XRCC1, on human chromosome 19. Cytogenet Cell Genet. 1989;52(1-2):11–14. [PubMed]
  • Myles GM, Sancar A. DNA repair. Chem Res Toxicol. 1989 Jul-Aug;2(4):197–226. [PubMed]
  • Okayama H, Berg P. A cDNA cloning vector that permits expression of cDNA inserts in mammalian cells. Mol Cell Biol. 1983 Feb;3(2):280–289. [PMC free article] [PubMed]
  • Pinkel D, Thompson LH, Gray JW, Vanderlaan M. Measurement of sister chromatid exchanges at very low bromodeoxyuridine substitution levels using a monoclonal antibody in Chinese hamster ovary cells. Cancer Res. 1985 Nov;45(11 Pt 2):5795–5798. [PubMed]
  • Potter H, Dressler D. A 'Southern Cross' method for the analysis of genome organization and the localization of transcription units. Gene. 1986;48(2-3):229–239. [PubMed]
  • Robson CN, Harris AL, Hickson ID. Isolation and characterization of Chinese hamster ovary cell lines sensitive to mitomycin C and bleomycin. Cancer Res. 1985 Nov;45(11 Pt 1):5304–5309. [PubMed]
  • Robson CN, Harris AL, Hickson ID. Defective repair of DNA single- and double-strand breaks in the bleomycin- and X-ray-sensitive Chinese hamster ovary cell mutant, BLM-2. Mutat Res. 1989 Mar;217(2):93–100. [PubMed]
  • Siciliano MJ, Carrano AV, Thompson LH. Assignment of a human DNA-repair gene associated with sister-chromatid exchange to chromosome 19. Mutat Res. 1986 Aug;174(4):303–308. [PubMed]
  • Southern EM. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. [PubMed]
  • Stamato TD, Weinstein R, Giaccia A, Mackenzie L. Isolation of cell cycle-dependent gamma ray-sensitive Chinese hamster ovary cell. Somatic Cell Genet. 1983 Mar;9(2):165–173. [PubMed]
  • Thompson LH. Somatic cell genetics approach to dissecting mammalian DNA repair. Environ Mol Mutagen. 1989;14(4):264–281. [PubMed]
  • Thompson LH, Bachinski LL, Stallings RL, Dolf G, Weber CA, Westerveld A, Siciliano MJ. Complementation of repair gene mutations on the hemizygous chromosome 9 in CHO: a third repair gene on human chromosome 19. Genomics. 1989 Nov;5(4):670–679. [PubMed]
  • Thompson LH, Brookman KW, Dillehay LE, Carrano AV, Mazrimas JA, Mooney CL, Minkler JL. A CHO-cell strain having hypersensitivity to mutagens, a defect in DNA strand-break repair, and an extraordinary baseline frequency of sister-chromatid exchange. Mutat Res. 1982 Aug;95(2-3):427–440. [PubMed]
  • Thompson LH, Brookman KW, Dillehay LE, Mooney CL, Carrano AV. Hypersensitivity to mutation and sister-chromatid-exchange induction in CHO cell mutants defective in incising DNA containing UV lesions. Somatic Cell Genet. 1982 Nov;8(6):759–773. [PubMed]
  • Thompson LH, Brookman KW, Minkler JL, Fuscoe JC, Henning KA, Carrano AV. DNA-mediated transfer of a human DNA repair gene that controls sister chromatid exchange. Mol Cell Biol. 1985 Apr;5(4):881–884. [PMC free article] [PubMed]
  • Thompson LH, Fong S, Brookman K. Validation of conditions for efficient detection of HPRT and APRT mutations in suspension-cultured Chinese hamster ovary cells. Mutat Res. 1980 Feb;74(1):21–36. [PubMed]
  • Thompson LH, Salazar EP, Brookman KW, Collins CC, Stewart SA, Busch DB, Weber CA. Recent progress with the DNA repair mutants of Chinese hamster ovary cells. J Cell Sci Suppl. 1987;6:97–110. [PubMed]
  • vanAnkeren SC, Murray D, Meyn RE. Induction and rejoining of gamma-ray-induced DNA single- and double-strand breaks in Chinese hamster AA8 cells and in two radiosensitive clones. Radiat Res. 1988 Dec;116(3):511–525. [PubMed]
  • van Duin M, de Wit J, Odijk H, Westerveld A, Yasui A, Koken MH, Hoeijmakers JH, Bootsma D. Molecular characterization of the human excision repair gene ERCC-1: cDNA cloning and amino acid homology with the yeast DNA repair gene RAD10. Cell. 1986 Mar 28;44(6):913–923. [PubMed]
  • Walker GC. Inducible DNA repair systems. Annu Rev Biochem. 1985;54:425–457. [PubMed]
  • Walker GC, Marsh L, Dodson LA. Genetic analyses of DNA repair: inference and extrapolation. Annu Rev Genet. 1985;19:103–126. [PubMed]
  • Weber CA, Salazar EP, Stewart SA, Thompson LH. Molecular cloning and biological characterization of a human gene, ERCC2, that corrects the nucleotide excision repair defect in CHO UV5 cells. Mol Cell Biol. 1988 Mar;8(3):1137–1146. [PMC free article] [PubMed]
  • Weber CA, Salazar EP, Stewart SA, Thompson LH. ERCC2: cDNA cloning and molecular characterization of a human nucleotide excision repair gene with high homology to yeast RAD3. EMBO J. 1990 May;9(5):1437–1447. [PubMed]
  • Weeda G, van Ham RC, Masurel R, Westerveld A, Odijk H, de Wit J, Bootsma D, van der Eb AJ, Hoeijmakers JH. Molecular cloning and biological characterization of the human excision repair gene ERCC-3. Mol Cell Biol. 1990 Jun;10(6):2570–2581. [PMC free article] [PubMed]
  • Westerveld A, Hoeijmakers JH, van Duin M, de Wit J, Odijk H, Pastink A, Wood RD, Bootsma D. Molecular cloning of a human DNA repair gene. Nature. 1984 Aug 2;310(5976):425–429. [PubMed]
  • Wood RD, Burki HJ. Repair capability and the cellular age response for killing and mutation induction after UV. Mutat Res. 1982 Aug;95(2-3):505–514. [PubMed]
  • Zdzienicka MZ, Tran Q, van der Schans GP, Simons JW. Characterization of an X-ray-hypersensitive mutant of V79 Chinese hamster cells. Mutat Res. 1988 Nov;194(3):239–249. [PubMed]

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