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
. 1993 Nov 1;90(21):9823–9827. doi: 10.1073/pnas.90.21.9823

Repair by human cell extracts of single (6-4) and cyclobutane thymine-thymine photoproducts in DNA.

D E Szymkowski 1, C W Lawrence 1, R D Wood 1
PMCID: PMC47664  PMID: 8234319

Abstract

One cis-syn cyclobutane thymine dimer or one (6-4) thymine-thymine photoproduct was built into an identical sequence of a closed-circular M13 duplex DNA, and nucleotide excision repair synthesis carried out by human cell extracts in the area containing each lesion was determined. Extracts from normal cells repaired the (6-4) photoproduct with a patch size of approximately 20-30 nucleotides, but repair was at least 10-fold lower at the cyclobutane dimer. The (6-4) lesion was repaired with comparable efficiency to a single acetylamino-fluorene-guanine adduct in a similar location. Extract from nucleotide excision repair-deficient xeroderma pigmentosum group A cells could not remove any of these adducts but could complete repair of the lesions after incision with Escherichia coli UvrABC proteins. This direct comparison of repair of two UV photoproducts, in an in vitro system where chromatin assembly and transcription are absent, suggests that the more rapid repair of the (6-4) lesion observed in the mammalian cell genome overall is due in part to a significant difference in the ability of the repair complex to locate and incise these lesions in DNA.

Full text

PDF
9823

Images in this article

9824Fig. 1
on p.9824
9825Fig. 2
on p.9825
9826Fig. 3
on p.9826
9826Fig. 4
on p.9826
9826Fig. 5
on p.9826

Selected References

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

  1. Banerjee S. K., Christensen R. B., Lawrence C. W., LeClerc J. E. Frequency and spectrum of mutations produced by a single cis-syn thymine-thymine cyclobutane dimer in a single-stranded vector. Proc Natl Acad Sci U S A. 1988 Nov;85(21):8141–8145. doi: 10.1073/pnas.85.21.8141. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Brash D. E., Seetharam S., Kraemer K. H., Seidman M. M., Bredberg A. Photoproduct frequency is not the major determinant of UV base substitution hot spots or cold spots in human cells. Proc Natl Acad Sci U S A. 1987 Jun;84(11):3782–3786. doi: 10.1073/pnas.84.11.3782. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Broughton B. C., Lehmann A. R., Harcourt S. A., Arlett C. F., Sarasin A., Kleijer W. J., Beemer F. A., Nairn R., Mitchell D. L. Relationship between pyrimidine dimers, 6-4 photoproducts, repair synthesis and cell survival: studies using cells from patients with trichothiodystrophy. Mutat Res. 1990 Jan;235(1):33–40. doi: 10.1016/0921-8777(90)90020-6. [DOI] [PubMed] [Google Scholar]
  4. Cleaver J. E., Cortés F., Karentz D., Lutze L. H., Morgan W. F., Player A. N., Vuksanovic L., Mitchell D. L. The relative biological importance of cyclobutane and (6-4) pyrimidine-pyrimidone dimer photoproducts in human cells: evidence from a xeroderma pigmentosum revertant. Photochem Photobiol. 1988 Jul;48(1):41–49. doi: 10.1111/j.1751-1097.1988.tb02784.x. [DOI] [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. Cleaver J. E., Jen J., Charles W. C., Mitchell D. L. Cyclobutane dimers and (6-4) photoproducts in human cells are mended with the same patch sizes. Photochem Photobiol. 1991 Sep;54(3):393–402. doi: 10.1111/j.1751-1097.1991.tb02033.x. [DOI] [PubMed] [Google Scholar]
  7. Franklin W. A., Doetsch P. W., Haseltine W. A. Structural determination of the ultraviolet light-induced thymine-cytosine pyrimidine-pyrimidone (6-4) photoproduct. Nucleic Acids Res. 1985 Jul 25;13(14):5317–5325. doi: 10.1093/nar/13.14.5317. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Franklin W. A., Haseltine W. A. Removal of UV light-induced pyrimidine-pyrimidone(6-4) products from Escherichia coli DNA requires the uvrA, uvrB, and urvC gene products. Proc Natl Acad Sci U S A. 1984 Jun;81(12):3821–3824. doi: 10.1073/pnas.81.12.3821. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gale J. M., Smerdon M. J. UV induced (6-4) photoproducts are distributed differently than cyclobutane dimers in nucleosomes. Photochem Photobiol. 1990 Apr;51(4):411–417. doi: 10.1111/j.1751-1097.1990.tb01732.x. [DOI] [PubMed] [Google Scholar]
  10. Hansson J., Grossman L., Lindahl T., Wood R. D. Complementation of the xeroderma pigmentosum DNA repair synthesis defect with Escherichia coli UvrABC proteins in a cell-free system. Nucleic Acids Res. 1990 Jan 11;18(1):35–40. doi: 10.1093/nar/18.1.35. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hansson J., Munn M., Rupp W. D., Kahn R., Wood R. D. Localization of DNA repair synthesis by human cell extracts to a short region at the site of a lesion. J Biol Chem. 1989 Dec 25;264(36):21788–21792. [PubMed] [Google Scholar]
  12. Huang J. C., Svoboda D. L., Reardon J. T., Sancar A. Human nucleotide excision nuclease removes thymine dimers from DNA by incising the 22nd phosphodiester bond 5' and the 6th phosphodiester bond 3' to the photodimer. Proc Natl Acad Sci U S A. 1992 Apr 15;89(8):3664–3668. doi: 10.1073/pnas.89.8.3664. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hutchinson F., Yamamoto K., Stein J., Wood R. D. Effect of photoreactivation on mutagenesis of lambda phage by ultraviolet light. J Mol Biol. 1988 Aug 5;202(3):593–601. doi: 10.1016/0022-2836(88)90289-6. [DOI] [PubMed] [Google Scholar]
  14. Hwang B. J., Chu G. Purification and characterization of a human protein that binds to damaged DNA. Biochemistry. 1993 Feb 16;32(6):1657–1666. doi: 10.1021/bi00057a033. [DOI] [PubMed] [Google Scholar]
  15. Koehl P., Burnouf D., Fuchs R. P. Construction of plasmids containing a unique acetylaminofluorene adduct located within a mutation hot spot. A new probe for frameshift mutagenesis. J Mol Biol. 1989 May 20;207(2):355–364. doi: 10.1016/0022-2836(89)90259-3. [DOI] [PubMed] [Google Scholar]
  16. LeClerc J. E., Borden A., Lawrence C. W. The thymine-thymine pyrimidine-pyrimidone(6-4) ultraviolet light photoproduct is highly mutagenic and specifically induces 3' thymine-to-cytosine transitions in Escherichia coli. Proc Natl Acad Sci U S A. 1991 Nov 1;88(21):9685–9689. doi: 10.1073/pnas.88.21.9685. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lommel L., Hanawalt P. C. Increased UV resistance of a xeroderma pigmentosum revertant cell line is correlated with selective repair of the transcribed strand of an expressed gene. Mol Cell Biol. 1993 Feb;13(2):970–976. doi: 10.1128/mcb.13.2.970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Meyn R. E., Vizard D. L., Hewitt R. R., Humphrey R. M. The fate of pyrimidine dimers in the DNA of ultraviolet-irradiated Chinese hamster cells. Photochem Photobiol. 1974 Sep;20(3):221–226. doi: 10.1111/j.1751-1097.1974.tb06570.x. [DOI] [PubMed] [Google Scholar]
  19. Mitchell D. L., Brash D. E., Nairn R. S. Rapid repair kinetics of pyrimidine(6-4)pyrimidone photoproducts in human cells are due to excision rather than conformational change. Nucleic Acids Res. 1990 Feb 25;18(4):963–971. doi: 10.1093/nar/18.4.963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Mitchell D. L., Clarkson J. M., Chao C. C., Rosenstein B. S. Repair of cyclobutane dimers and (6-4) photoproducts in ICR 2A frog cells. Photochem Photobiol. 1986 May;43(5):595–597. doi: 10.1111/j.1751-1097.1986.tb09539.x. [DOI] [PubMed] [Google Scholar]
  21. Mitchell D. L., Haipek C. A., Clarkson J. M. (6-4)Photoproducts are removed from the DNA of UV-irradiated mammalian cells more efficiently than cyclobutane pyrimidine dimers. Mutat Res. 1985 Jul;143(3):109–112. doi: 10.1016/s0165-7992(85)80018-x. [DOI] [PubMed] [Google Scholar]
  22. Mitchell D. L., Nairn R. S. The biology of the (6-4) photoproduct. Photochem Photobiol. 1989 Jun;49(6):805–819. doi: 10.1111/j.1751-1097.1989.tb05578.x. [DOI] [PubMed] [Google Scholar]
  23. Mitchell D. L., Nguyen T. D., Cleaver J. E. Nonrandom induction of pyrimidine-pyrimidone (6-4) photoproducts in ultraviolet-irradiated human chromatin. J Biol Chem. 1990 Apr 5;265(10):5353–5356. [PubMed] [Google Scholar]
  24. Myles G. M., Van Houten B., Sancar A. Utilization of DNA photolyase, pyrimidine dimer endonucleases, and alkali hydrolysis in the analysis of aberrant ABC excinuclease incisions adjacent to UV-induced DNA photoproducts. Nucleic Acids Res. 1987 Feb 11;15(3):1227–1243. doi: 10.1093/nar/15.3.1227. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Rao S. N., Kollman P. A. Conformations of deoxydodecanucleotides with pyrimidine (6-4)-pyrimidone photoadducts. Photochem Photobiol. 1985 Nov;42(5):465–475. doi: 10.1111/j.1751-1097.1985.tb01596.x. [DOI] [PubMed] [Google Scholar]
  26. Rinaldy A., Bellew T., Egli E., Lloyd R. S. Increased UV resistance in xeroderma pigmentosum group A cells after transformation with a human genomic DNA clone. Proc Natl Acad Sci U S A. 1990 Sep;87(17):6818–6822. doi: 10.1073/pnas.87.17.6818. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Robins P., Jones C. J., Biggerstaff M., Lindahl T., Wood R. D. Complementation of DNA repair in xeroderma pigmentosum group A cell extracts by a protein with affinity for damaged DNA. EMBO J. 1991 Dec;10(12):3913–3921. doi: 10.1002/j.1460-2075.1991.tb04961.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Sancar A., Franklin K. A., Sancar G., Tang M. S. Repair of psoralen and acetylaminofluorene DNA adducts by ABC excinuclease. J Mol Biol. 1985 Aug 20;184(4):725–734. doi: 10.1016/0022-2836(85)90316-x. [DOI] [PubMed] [Google Scholar]
  29. Sancar A., Rupp W. D. A novel repair enzyme: UVRABC excision nuclease of Escherichia coli cuts a DNA strand on both sides of the damaged region. Cell. 1983 May;33(1):249–260. doi: 10.1016/0092-8674(83)90354-9. [DOI] [PubMed] [Google Scholar]
  30. Shivji K. K., Kenny M. K., Wood R. D. Proliferating cell nuclear antigen is required for DNA excision repair. Cell. 1992 Apr 17;69(2):367–374. doi: 10.1016/0092-8674(92)90416-a. [DOI] [PubMed] [Google Scholar]
  31. Spivak G., Hanawalt P. C. Translesion DNA synthesis in the dihydrofolate reductase domain of UV-irradiated CHO cells. Biochemistry. 1992 Jul 28;31(29):6794–6800. doi: 10.1021/bi00144a021. [DOI] [PubMed] [Google Scholar]
  32. Szymkowski D. E., Hajibagheri M. A., Wood R. D. Electron microscopy of DNA excision repair patches produced by human cell extracts. J Mol Biol. 1993 May 20;231(2):251–260. doi: 10.1006/jmbi.1993.1280. [DOI] [PubMed] [Google Scholar]
  33. Szymkowski D. E., Yarema K., Essigmann J. M., Lippard S. J., Wood R. D. An intrastrand d(GpG) platinum crosslink in duplex M13 DNA is refractory to repair by human cell extracts. Proc Natl Acad Sci U S A. 1992 Nov 15;89(22):10772–10776. doi: 10.1073/pnas.89.22.10772. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Thomas D. C., Husain I., Chaney S. G., Panigrahi G. B., Walker I. G. Sequence effect on incision by (A)BC excinuclease of 4NQO adducts and UV photoproducts. Nucleic Acids Res. 1991 Jan 25;19(2):365–370. doi: 10.1093/nar/19.2.365. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Thompson L. H., Mitchell D. L., Regan J. D., Bouffler S. D., Stewart S. A., Carrier W. L., Nairn R. S., Johnson R. T. CHO mutant UV61 removes (6-4) photoproducts but not cyclobutane dimers. Mutagenesis. 1989 Mar;4(2):140–146. doi: 10.1093/mutage/4.2.140. [DOI] [PubMed] [Google Scholar]
  36. Varghese A. J., Wang S. Y. Thymine-thymine adduct as a photoproduct of thymine. Science. 1968 Apr 12;160(3824):186–187. doi: 10.1126/science.160.3824.186. [DOI] [PubMed] [Google Scholar]
  37. Wood R. D. Repair of pyrimidine dimer ultraviolet light photoproducts by human cell extracts. Biochemistry. 1989 Oct 17;28(21):8287–8292. doi: 10.1021/bi00447a005. [DOI] [PubMed] [Google Scholar]
  38. Wood R. D., Robins P. DNA repair replication by soluble extracts from human lymphoid cell lines. Genome. 1989;31(2):601–604. doi: 10.1139/g89-112. [DOI] [PubMed] [Google Scholar]
  39. 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]
  40. Zdzienicka M. Z., Venema J., Mitchell D. L., van Hoffen A., van Zeeland A. A., Vrieling H., Mullenders L. H., Lohman P. H., Simons J. W. (6-4) photoproducts and not cyclobutane pyrimidine dimers are the main UV-induced mutagenic lesions in Chinese hamster cells. Mutat Res. 1992 Jan;273(1):73–83. doi: 10.1016/0921-8777(92)90051-4. [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