Skip to main content
PMC home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1993 Dec 25;21(25):5890–5895. doi: 10.1093/nar/21.25.5890

Deficient repair of the transcribed strand of active genes in Cockayne's syndrome cells.

A van Hoffen 1, A T Natarajan 1, L V Mayne 1, A A van Zeeland 1, L H Mullenders 1, J Venema 1
PMCID: PMC310470  PMID: 8290349

Abstract

Removal of ultraviolet light induced cyclobutane pyrimidine dimers (CPD) from active and inactive genes was analyzed in cells derived from patients suffering from the hereditary disease Cockayne's syndrome (CS) using strand specific probes. The results indicate that the defect in CS cells affects two levels of repair of lesions in active genes. Firstly, CS cells are deficient in selective repair of the transcribed strand of active genes. In these cells the rate and efficiency of repair of CPD are equal for the transcribed and the nontranscribed strand of the active ADA and DHFR genes. In normal cells on the other hand, the transcribed strand of these genes is repaired faster than the nontranscribed strand. However, the nontranscribed strand is still repaired more efficiently than the inactive 754 gene and the gene coding for coagulation factor IX. Secondly, the repair level of active genes in CS cells exceeds that of inactive loci but is slower than the nontranscribed strand of active genes in normal cells. Our results support the model that CS cells lack a factor which is involved in targeting repair enzymes specifically towards DNA damage located in (potentially) active DNA.

Full text

PDF
5895

Images in this article

5892Image
on p.5892
5892Image
on p.5892

Selected References

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

  1. Anson D. S., Choo K. H., Rees D. J., Giannelli F., Gould K., Huddleston J. A., Brownlee G. G. The gene structure of human anti-haemophilic factor IX. EMBO J. 1984 May;3(5):1053–1060. doi: 10.1002/j.1460-2075.1984.tb01926.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Antonarakis S. E. The molecular genetics of hemophilia A and B in man. Factor VIII and factor IX deficiency. Adv Hum Genet. 1988;17:27–59. doi: 10.1007/978-1-4613-0987-1_2. [DOI] [PubMed] [Google Scholar]
  3. Arlett C. F., Green M. H., Priestley A., Harcourt S. A., Mayne L. V. Comparative human cellular radiosensitivity: I. The effect of SV40 transformation and immortalisation on the gamma-irradiation survival of skin derived fibroblasts from normal individuals and from ataxia-telangiectasia patients and heterozygotes. Int J Radiat Biol. 1988 Dec;54(6):911–928. doi: 10.1080/09553008814552321. [DOI] [PubMed] [Google Scholar]
  4. Berkvens T. M., Gerritsen E. J., Oldenburg M., Breukel C., Wijnen J. T., van Ormondt H., Vossen J. M., van der Eb A. J., Meera Khan P. Severe combined immune deficiency due to a homozygous 3.2-kb deletion spanning the promoter and first exon of the adenosine deaminase gene. Nucleic Acids Res. 1987 Nov 25;15(22):9365–9378. doi: 10.1093/nar/15.22.9365. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Biernat J., Göbel U. B., Köster H. New bacteriophage vectors for the large scale production of single stranded insert DNA. J Biochem Biophys Methods. 1989 Aug-Sep;19(2-3):155–167. doi: 10.1016/0165-022x(89)90023-7. [DOI] [PubMed] [Google Scholar]
  6. Bohr V. A., Smith C. A., Okumoto D. S., Hanawalt P. C. DNA repair in an active gene: removal of pyrimidine dimers from the DHFR gene of CHO cells is much more efficient than in the genome overall. Cell. 1985 Feb;40(2):359–369. doi: 10.1016/0092-8674(85)90150-3. [DOI] [PubMed] [Google Scholar]
  7. Elliott G. C., Downes C. S. Qualitative differences between replicative and repair synthesis of DNA in normal and transformed mouse cells as measured by precursor discrimination. Mutat Res. 1986 Nov;166(3):295–302. doi: 10.1016/0167-8817(86)90029-5. [DOI] [PubMed] [Google Scholar]
  8. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  9. Hofker M. H., van Ommen G. J., Bakker E., Burmeister M., Pearson P. L. Development of additional RFLP probes near the locus for Duchenne muscular dystrophy by cosmid cloning of the DXS84 (754) locus. Hum Genet. 1986 Nov;74(3):270–274. doi: 10.1007/BF00282547. [DOI] [PubMed] [Google Scholar]
  10. Lattier D. L., States J. C., Hutton J. J., Wiginton D. A. Cell type-specific transcriptional regulation of the human adenosine deaminase gene. Nucleic Acids Res. 1989 Feb 11;17(3):1061–1076. doi: 10.1093/nar/17.3.1061. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Madhani H. D., Bohr V. A., Hanawalt P. C. Differential DNA repair in transcriptionally active and inactive proto-oncogenes: c-abl and c-mos. Cell. 1986 May 9;45(3):417–423. doi: 10.1016/0092-8674(86)90327-2. [DOI] [PubMed] [Google Scholar]
  12. Mayne L. V., Lehmann A. R. Failure of RNA synthesis to recover after UV irradiation: an early defect in cells from individuals with Cockayne's syndrome and xeroderma pigmentosum. Cancer Res. 1982 Apr;42(4):1473–1478. [PubMed] [Google Scholar]
  13. Mayne L. V., Lehmann A. R., Waters R. Excision repair in Cockayne syndrome. Mutat Res. 1982 Nov;106(1):179–189. doi: 10.1016/0027-5107(82)90200-7. [DOI] [PubMed] [Google Scholar]
  14. Mellon I., Bohr V. A., Smith C. A., Hanawalt P. C. Preferential DNA repair of an active gene in human cells. Proc Natl Acad Sci U S A. 1986 Dec;83(23):8878–8882. doi: 10.1073/pnas.83.23.8878. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Mellon I., Hanawalt P. C. Induction of the Escherichia coli lactose operon selectively increases repair of its transcribed DNA strand. Nature. 1989 Nov 2;342(6245):95–98. doi: 10.1038/342095a0. [DOI] [PubMed] [Google Scholar]
  16. Mellon I., Spivak G., Hanawalt P. C. Selective removal of transcription-blocking DNA damage from the transcribed strand of the mammalian DHFR gene. Cell. 1987 Oct 23;51(2):241–249. doi: 10.1016/0092-8674(87)90151-6. [DOI] [PubMed] [Google Scholar]
  17. Mullenders L. H., Vrieling H., Venema J., van Zeeland A. A. Hierarchies of DNA repair in mammalian cells: biological consequences. Mutat Res. 1991 Sep-Oct;250(1-2):223–228. doi: 10.1016/0027-5107(91)90179-r. [DOI] [PubMed] [Google Scholar]
  18. Mullenders L. H., van Kesteren van Leeuwen A. C., van Zeeland A. A., Natarajan A. T. Nuclear matrix associated DNA is preferentially repaired in normal human fibroblasts, exposed to a low dose of ultraviolet light but not in Cockayne's syndrome fibroblasts. Nucleic Acids Res. 1988 Nov 25;16(22):10607–10622. doi: 10.1093/nar/16.22.10607. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Nakabeppu Y., Yamashita K., Sekiguchi M. Purification and characterization of normal and mutant forms of T4 endonuclease V. J Biol Chem. 1982 Mar 10;257(5):2556–2562. [PubMed] [Google Scholar]
  20. Palmer T. D., Thompson A. R., Miller A. D. Production of human factor IX in animals by genetically modified skin fibroblasts: potential therapy for hemophilia B. Blood. 1989 Feb;73(2):438–445. [PubMed] [Google Scholar]
  21. Santiago C., Collins M., Johnson L. F. In vitro and in vivo analysis of the control of dihydrofolate reductase gene transcription in serum-stimulated mouse fibroblasts. J Cell Physiol. 1984 Jan;118(1):79–86. doi: 10.1002/jcp.1041180114. [DOI] [PubMed] [Google Scholar]
  22. Selby C. P., Sancar A. Gene- and strand-specific repair in vitro: partial purification of a transcription-repair coupling factor. Proc Natl Acad Sci U S A. 1991 Sep 15;88(18):8232–8236. doi: 10.1073/pnas.88.18.8232. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Selby C. P., Witkin E. M., Sancar A. Escherichia coli mfd mutant deficient in "mutation frequency decline" lacks strand-specific repair: in vitro complementation with purified coupling factor. Proc Natl Acad Sci U S A. 1991 Dec 15;88(24):11574–11578. doi: 10.1073/pnas.88.24.11574. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Sweder K. S., Hanawalt P. C. Preferential repair of cyclobutane pyrimidine dimers in the transcribed strand of a gene in yeast chromosomes and plasmids is dependent on transcription. Proc Natl Acad Sci U S A. 1992 Nov 15;89(22):10696–10700. doi: 10.1073/pnas.89.22.10696. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Troelstra C., van Gool A., de Wit J., Vermeulen W., Bootsma D., Hoeijmakers J. H. ERCC6, a member of a subfamily of putative helicases, is involved in Cockayne's syndrome and preferential repair of active genes. Cell. 1992 Dec 11;71(6):939–953. doi: 10.1016/0092-8674(92)90390-x. [DOI] [PubMed] [Google Scholar]
  26. Venema J., Bartosová Z., Natarajan A. T., van Zeeland A. A., Mullenders L. H. Transcription affects the rate but not the extent of repair of cyclobutane pyrimidine dimers in the human adenosine deaminase gene. J Biol Chem. 1992 May 5;267(13):8852–8856. [PubMed] [Google Scholar]
  27. Venema J., Mullenders L. H., Natarajan A. T., van Zeeland A. A., Mayne L. V. The genetic defect in Cockayne syndrome is associated with a defect in repair of UV-induced DNA damage in transcriptionally active DNA. Proc Natl Acad Sci U S A. 1990 Jun;87(12):4707–4711. doi: 10.1073/pnas.87.12.4707. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Venema J., van Hoffen A., Karcagi V., Natarajan A. T., van Zeeland A. A., Mullenders L. H. Xeroderma pigmentosum complementation group C cells remove pyrimidine dimers selectively from the transcribed strand of active genes. Mol Cell Biol. 1991 Aug;11(8):4128–4134. doi: 10.1128/mcb.11.8.4128. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Venema J., van Hoffen A., Natarajan A. T., van Zeeland A. A., Mullenders L. H. The residual repair capacity of xeroderma pigmentosum complementation group C fibroblasts is highly specific for transcriptionally active DNA. Nucleic Acids Res. 1990 Feb 11;18(3):443–448. doi: 10.1093/nar/18.3.443. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Will C. L., Dolnick B. J. 5-Fluorouracil augmentation of dihydrofolate reductase gene transcripts containing intervening sequences in methotrexate-resistant KB cells. Mol Pharmacol. 1986 Jun;29(6):643–648. [PubMed] [Google Scholar]
  31. Yoshitake S., Schach B. G., Foster D. C., Davie E. W., Kurachi K. Nucleotide sequence of the gene for human factor IX (antihemophilic factor B). Biochemistry. 1985 Jul 2;24(14):3736–3750. doi: 10.1021/bi00335a049. [DOI] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES