Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1989 Sep;86(18):6997–7001. doi: 10.1073/pnas.86.18.6997

Molecular cloning and characterization of a mammalian excision repair gene that partially restores UV resistance to xeroderma pigmentosum complementation group D cells.

J E Arrand 1, N M Bone 1, R T Johnson 1
PMCID: PMC297979  PMID: 2780557

Abstract

A hamster DNA repair gene has been isolated by cosmid rescue after two rounds of transfection of an immortalized xeroderma pigmentosum (XP) complementation group D cell line with neomycin-resistance gene (neo)-tagged normal hamster DNA and selection with G418 and ultraviolet irradiation. The functional length of the sequence has been defined as 11.5 kilobase pairs by measurement of the region of overlap between two hamster DNA-containing cosmids, cloned by selection for the integrated neo gene, that are able to confer an increase in resistance to ultraviolet irradiation on two XP-D cell line but not on an XP-A line. Detailed molecular characterization of the hamster repair gene has revealed no obvious similarities to two human excision repair genes (ERCC1 and ERCC2) that correct repair-defective hamster cells but have no effect on XP cells. Hybridization analyses of normal human and XP cell genomic DNAs and mRNAs, using a cosmid-clone probe from which repeated sequences have been removed, show that homologues are present and expressed in all cases.

Full text

PDF
6997

Images in this article

6999Image
on p.6999
7000Image
on p.7000

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Arrand J. E., Squires S., Bone N. M., Johnson R. T. Restoration of u.v.-induced excision repair in Xeroderma D cells transfected with the denV gene of bacteriophage T4. EMBO J. 1987 Oct;6(10):3125–3131. doi: 10.1002/j.1460-2075.1987.tb02622.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Brady G., Funk A., Mattern J., Schütz G., Brown R. Use of gene transfer and a novel cosmid rescue strategy to isolate transforming sequences. EMBO J. 1985 Oct;4(10):2583–2588. doi: 10.1002/j.1460-2075.1985.tb03974.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Canaani D., Naiman T., Teitz T., Berg P. Immortalization of xeroderma pigmentosum cells by simian virus 40 DNA having a defective origin of DNA replication. Somat Cell Mol Genet. 1986 Jan;12(1):13–20. doi: 10.1007/BF01560723. [DOI] [PubMed] [Google Scholar]
  4. Chen C., Okayama H. High-efficiency transformation of mammalian cells by plasmid DNA. Mol Cell Biol. 1987 Aug;7(8):2745–2752. doi: 10.1128/mcb.7.8.2745. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cleaver J. E., Cortés F., Lutze L. H., Morgan W. F., Player A. N., Mitchell D. L. Unique DNA repair properties of a xeroderma pigmentosum revertant. Mol Cell Biol. 1987 Sep;7(9):3353–3357. doi: 10.1128/mcb.7.9.3353. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Collins A. R., Johnson R. T. Repair and survival after UV in quiescent and proliferating Microtus agrestis cells: different rates of incision and different dependence on DNA precursor supply. J Cell Physiol. 1979 Apr;99(1):125–137. doi: 10.1002/jcp.1040990114. [DOI] [PubMed] [Google Scholar]
  7. Corsaro C. M., Pearson M. L. Enhancing the efficiency of DNA-mediated gene transfer in mammalian cells. Somatic Cell Genet. 1981 Sep;7(5):603–616. doi: 10.1007/BF01549662. [DOI] [PubMed] [Google Scholar]
  8. Erixon K., Ahnström G. Single-strand breaks in DNA during repair of UV-induced damage in normal human and xeroderma pigmentosum cells as determined by alkaline DNA unwinding and hydroxylapatite chromatography: effects of hydroxyurea, 5-fluorodeoxyuridine and 1-beta-D-arabinofuranosylcytosine on the kinetics of repair. Mutat Res. 1979 Feb;59(2):257–271. doi: 10.1016/0027-5107(79)90164-7. [DOI] [PubMed] [Google Scholar]
  9. Feinberg A. P., Vogelstein B. "A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity". Addendum. Anal Biochem. 1984 Feb;137(1):266–267. doi: 10.1016/0003-2697(84)90381-6. [DOI] [PubMed] [Google Scholar]
  10. Graham F. L., van der Eb A. J. A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology. 1973 Apr;52(2):456–467. doi: 10.1016/0042-6822(73)90341-3. [DOI] [PubMed] [Google Scholar]
  11. Hoeijmakers J. H., Odijk H., Westerveld A. Differences between rodent and human cell lines in the amount of integrated DNA after transfection. Exp Cell Res. 1987 Mar;169(1):111–119. doi: 10.1016/0014-4827(87)90230-8. [DOI] [PubMed] [Google Scholar]
  12. Johnson R. T., Squires S., Elliott G. C., Koch G. L., Rainbow A. J. Xeroderma pigmentosum D-HeLa hybrids with low and high ultraviolet sensitivity associated with normal and diminished DNA repair ability, respectively. J Cell Sci. 1985 Jun;76:115–133. doi: 10.1242/jcs.76.1.115. [DOI] [PubMed] [Google Scholar]
  13. Mayne L. V., Jones T., Dean S. W., Harcourt S. A., Lowe J. E., Priestley A., Steingrimsdottir H., Sykes H., Green M. H., Lehmann A. R. SV 40-transformed normal and DNA-repair-deficient human fibroblasts can be transfected with high frequency but retain only limited amounts of integrated DNA. Gene. 1988 Jun 15;66(1):65–76. doi: 10.1016/0378-1119(88)90225-9. [DOI] [PubMed] [Google Scholar]
  14. Paterson M. C., Gentner N. E., Middlestadt M. V., Weinfeld M. Cancer predisposition, carcinogen hypersensitivity, and aberrant DNA metabolism. J Cell Physiol Suppl. 1984;3:45–62. doi: 10.1002/jcp.1041210408. [DOI] [PubMed] [Google Scholar]
  15. 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. doi: 10.1016/0378-1119(86)90081-8. [DOI] [PubMed] [Google Scholar]
  16. Protić-Sabljić M., Seetharam S., Seidman M. M., Kraemer K. H. An SV40-transformed xeroderma pigmentosum group D cell line: establishment, ultraviolet sensitivity, transfection efficiency and plasmid mutation induction. Mutat Res. 1986 Nov;166(3):287–294. doi: 10.1016/0167-8817(86)90028-3. [DOI] [PubMed] [Google Scholar]
  17. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  18. Royer-Pokora B., Peterson W. D., Jr, Haseltine W. A. Biological and biochemical characterization of an SV40-transformed xeroderma pigmentosum cell line. Exp Cell Res. 1984 Apr;151(2):408–420. doi: 10.1016/0014-4827(84)90391-4. [DOI] [PubMed] [Google Scholar]
  19. Sealey P. G., Whittaker P. A., Southern E. M. Removal of repeated sequences from hybridisation probes. Nucleic Acids Res. 1985 Mar 25;13(6):1905–1922. doi: 10.1093/nar/13.6.1905. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  21. Squires S., Johnson R. T., Collins A. R. Initial rates of DNA incision in UV-irradiated human cells: differences between normal, xeroderma pigmentosum and tumour cells. Mutat Res. 1982 Aug;95(2-3):389–404. doi: 10.1016/0027-5107(82)90273-1. [DOI] [PubMed] [Google Scholar]
  22. Takebe H., Nii S., Ishii M. I., Utsumi H. Comparative studies of host-cell reactivation, colony forming ability and excision repair after UV irradiation of xeroderma pigmentosum, normal human and some other mammalian cells. Mutat Res. 1974 Dec;25(3):383–390. doi: 10.1016/0027-5107(74)90067-0. [DOI] [PubMed] [Google Scholar]
  23. Teitz T., Naiman T., Avissar S. S., Bar S., Okayama H., Canaani D. Complementation of the UV-sensitive phenotype of a xeroderma pigmentosum human cell line by transfection with a cDNA clone library. Proc Natl Acad Sci U S A. 1987 Dec;84(24):8801–8804. doi: 10.1073/pnas.84.24.8801. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Thompson L. H., Mooney C. L., Brookman K. W. Genetic complementation between UV-sensitive CHO mutants and xeroderma pigmentosum fibroblasts. Mutat Res. 1985 Jun-Jul;150(1-2):423–429. doi: 10.1016/0027-5107(85)90139-3. [DOI] [PubMed] [Google Scholar]
  25. Weber C. A., Salazar E. P., Stewart S. A., Thompson L. H. 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. doi: 10.1128/mcb.8.3.1137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Westerveld A., Hoeijmakers J. H., van Duin M., de Wit J., Odijk H., Pastink A., Wood R. D., Bootsma D. Molecular cloning of a human DNA repair gene. Nature. 1984 Aug 2;310(5976):425–429. doi: 10.1038/310425a0. [DOI] [PubMed] [Google Scholar]
  27. Wood R. D., Robins P., Lindahl T. Complementation of the xeroderma pigmentosum DNA repair defect in cell-free extracts. Cell. 1988 Apr 8;53(1):97–106. doi: 10.1016/0092-8674(88)90491-6. [DOI] [PubMed] [Google Scholar]
  28. van Duin M., Janssen J. H., de Wit J., Hoeijmakers J. H., Thompson L. H., Bootsma D., Westerveld A. Transfection of the cloned human excision repair gene ERCC-1 to UV-sensitive CHO mutants only corrects the repair defect in complementation group-2 mutants. Mutat Res. 1988 Mar;193(2):123–130. doi: 10.1016/0167-8817(88)90042-9. [DOI] [PubMed] [Google Scholar]
  29. van Duin M., Koken M. H., van den Tol J., ten Dijke P., Odijk H., Westerveld A., Bootsma D., Hoeijmakers J. H. Genomic characterization of the human DNA excision repair gene ERCC-1. Nucleic Acids Res. 1987 Nov 25;15(22):9195–9213. doi: 10.1093/nar/15.22.9195. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. van Duin M., Vredeveldt G., Mayne L. V., Odijk H., Vermeulen W., Klein B., Weeda G., Hoeijmakers J. H., Bootsma D., Westerveld A. The cloned human DNA excision repair gene ERCC-1 fails to correct xeroderma pigmentosum complementation groups A through I. Mutat Res. 1989 Mar;217(2):83–92. doi: 10.1016/0921-8777(89)90059-1. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES