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. 1990 Sep;87(17):6818–6822. doi: 10.1073/pnas.87.17.6818

Increased UV resistance in xeroderma pigmentosum group A cells after transformation with a human genomic DNA clone.

A Rinaldy 1, T Bellew 1, E Egli 1, R S Lloyd 1
PMCID: PMC54629  PMID: 2168562

Abstract

Xeroderma pigmentosum (XP) is an autosomal recessive disease in which the major clinical manifestation is a 2,000-fold enhanced probability of developing sunlight-induced skin tumors, and the molecular basis for the disease is a defective DNA excision repair system. To clone the gene defective in XP complementation group A (XP-A), cDNA clones were isolated by a competition hybridization strategy in which the corresponding mRNAs were more abundant in cells of the obligately heterozygous parents relative to cells of the homozygous proband affected with the disease. In this report, a human genomic DNA clone that contains this cDNA was transformed into two independent homozygous XP-A cell lines, and these transformants displayed partial restoration of resistance to the killing effects of UV irradiation. The abundance of mRNA corresponding to this cDNA appears to correlate well with the observed UV cell survival. The results of unscheduled DNA synthesis after UV exposure indicate that the transformed cells are repair proficient relative to that of the control XP-A cells. However, using this same genomic DNA, transformation of an XP-F cell line did not confer any enhancement of UV survival or promote unscheduled DNA synthesis after UV exposure.

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Selected References

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

  1. Andrews A. D., Barrett S. F., Robbins J. H. Xeroderma pigmentosum neurological abnormalities correlate with colony-forming ability after ultraviolet radiation. Proc Natl Acad Sci U S A. 1978 Apr;75(4):1984–1988. doi: 10.1073/pnas.75.4.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Arrand J. E., Bone N. M., Johnson R. T. Molecular cloning and characterization of a mammalian excision repair gene that partially restores UV resistance to xeroderma pigmentosum complementation group D cells. Proc Natl Acad Sci U S A. 1989 Sep;86(18):6997–7001. doi: 10.1073/pnas.86.18.6997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cleaver J. E., Bootsma D. Xeroderma pigmentosum: biochemical and genetic characteristics. Annu Rev Genet. 1975;9:19–38. doi: 10.1146/annurev.ge.09.120175.000315. [DOI] [PubMed] [Google Scholar]
  4. Cleaver J. E., Karentz D. DNA repair in man: regulation by a multigene family and association with human disease. Bioessays. 1987 Mar;6(3):122–127. doi: 10.1002/bies.950060307. [DOI] [PubMed] [Google Scholar]
  5. Grunstein M., Hogness D. S. Colony hybridization: a method for the isolation of cloned DNAs that contain a specific gene. Proc Natl Acad Sci U S A. 1975 Oct;72(10):3961–3965. doi: 10.1073/pnas.72.10.3961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Lau Y. F., Kan Y. W. Versatile cosmid vectors for the isolation, expression, and rescue of gene sequences: studies with the human alpha-globin gene cluster. Proc Natl Acad Sci U S A. 1983 Sep;80(17):5225–5229. doi: 10.1073/pnas.80.17.5225. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Legerski R. J., Brown D. B., Peterson C. A., Robberson D. L. Transient complementation of xeroderma pigmentosum cells by microinjection of poly(A)+ RNA. Proc Natl Acad Sci U S A. 1984 Sep;81(18):5676–5679. doi: 10.1073/pnas.81.18.5676. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Maher V. M., Birch N., Otto J. R., MacCormick J. J. Cytotoxicity of carcinogenic aromatic amides in normal and xeroderma pigmentosum fibroblasts with different DNA repair capabilities. J Natl Cancer Inst. 1975 Jun;54(6):1287–1294. doi: 10.1093/jnci/54.6.1287. [DOI] [PubMed] [Google Scholar]
  9. Painter R. B. The action of ultraviolet light on mammalian cells. Photophysiology. 1970;5:169–189. [PubMed] [Google Scholar]
  10. Rinaldy A., Dodson M. L., Darling T. L., Lloyd R. S. Gene cloning using cDNA libraries in a differential competition hybridization strategy: application to cloning XP-A related genes. DNA. 1988 Oct;7(8):563–570. doi: 10.1089/dna.1.1988.7.563. [DOI] [PubMed] [Google Scholar]
  11. Robbins J. H., Kraemer K. H., Lutzner M. A., Festoff B. W., Coon H. G. Xeroderma pigmentosum. An inherited diseases with sun sensitivity, multiple cutaneous neoplasms, and abnormal DNA repair. Ann Intern Med. 1974 Feb;80(2):221–248. doi: 10.7326/0003-4819-80-2-221. [DOI] [PubMed] [Google Scholar]
  12. Setlow R. B. Repair deficient human disorders and cancer. Nature. 1978 Feb 23;271(5647):713–717. doi: 10.1038/271713a0. [DOI] [PubMed] [Google Scholar]
  13. Stefanini M., Keijzer W., Westerveld A., Bootsma D. Interspecies complementation analysis of xeroderma pigmentosum and UV-sensitive Chinese hamster cells. Exp Cell Res. 1985 Dec;161(2):373–380. doi: 10.1016/0014-4827(85)90094-1. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. Tanaka K., Satokata I., Ogita Z., Uchida T., Okada Y. Molecular cloning of a mouse DNA repair gene that complements the defect of group-A xeroderma pigmentosum. Proc Natl Acad Sci U S A. 1989 Jul;86(14):5512–5516. doi: 10.1073/pnas.86.14.5512. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Thompson L. H., Busch D. B., Brookman K., Mooney C. L., Glaser D. A. Genetic diversity of UV-sensitive DNA repair mutants of Chinese hamster ovary cells. Proc Natl Acad Sci U S A. 1981 Jun;78(6):3734–3737. doi: 10.1073/pnas.78.6.3734. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. 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]
  18. Thompson L. H., Mooney C. L., Burkhart-Schultz K., Carrano A. V., Siciliano M. J. Correction of a nucleotide-excision-repair mutation by human chromosome 19 in hamster-human hybrid cells. Somat Cell Mol Genet. 1985 Jan;11(1):87–92. doi: 10.1007/BF01534738. [DOI] [PubMed] [Google Scholar]
  19. Wigler M., Pellicer A., Silverstein S., Axel R., Urlaub G., Chasin L. DNA-mediated transfer of the adenine phosphoribosyltransferase locus into mammalian cells. Proc Natl Acad Sci U S A. 1979 Mar;76(3):1373–1376. doi: 10.1073/pnas.76.3.1373. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Yagi T., Takebe H. Establishment by SV40 transformation and characteristics of a cell line of xeroderma pigmentosum belonging to complementation group F. Mutat Res. 1983 Feb;112(1):59–66. doi: 10.1016/0167-8817(83)90024-x. [DOI] [PubMed] [Google Scholar]
  21. 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]

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