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. 1987 Dec;7(12):4317–4323. doi: 10.1128/mcb.7.12.4317

Repair of UV-induced lesions in Xenopus laevis oocytes.

R J Legerski 1, J E Penkala 1, C A Peterson 1, D A Wright 1
PMCID: PMC368114  PMID: 2830488

Abstract

We characterized a DNA repair system in frog oocytes by comicroinjection of UV-irradiated pBR322 DNA and radiolabeled nucleotides. Repair synthesis was monitored by incorporation of label into recovered pBR322 DNA and by a novel method in which the removal of UV photoproducts was determined from the shift of DNA topoisomers that occurs during gel electrophoresis upon repair of these lesions. We investigated the effects of several drugs in the oocyte system and found that although novobiocin, an inhibitor of topoisomerase II, was an effective inhibitor of repair, VM-26, another inhibitor of topoisomerase II, was not. In addition, the topoisomerase I inhibitor camptothecin had no effect on repair in this system. Finally, circular DNA (either supercoiled or nicked circular) was repaired at least 50 times more rapidly than linear DNA.

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Selected References

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

  1. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bohr V. A., Hanawalt P. C. Novobiocin does not inhibit DNA repair in an active gene. Carcinogenesis. 1986 Nov;7(11):1917–1920. doi: 10.1093/carcin/7.11.1917. [DOI] [PubMed] [Google Scholar]
  3. Carroll D. Genetic recombination of bacteriophage lambda DNAs in Xenopus oocytes. Proc Natl Acad Sci U S A. 1983 Nov;80(22):6902–6906. doi: 10.1073/pnas.80.22.6902. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chen G. L., Yang L., Rowe T. C., Halligan B. D., Tewey K. M., Liu L. F. Nonintercalative antitumor drugs interfere with the breakage-reunion reaction of mammalian DNA topoisomerase II. J Biol Chem. 1984 Nov 10;259(21):13560–13566. [PubMed] [Google Scholar]
  5. Collins A., Johnson R. Novobiocin; an inhibitor of the repair of UV-induced but not X-ray-induced damage in mammalian cells. Nucleic Acids Res. 1979 Nov 10;7(5):1311–1320. doi: 10.1093/nar/7.5.1311. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cook P. R., Brazell I. A. Detection and repair of single-strand breaks in nuclear DNA. Nature. 1976 Oct 21;263(5579):679–682. doi: 10.1038/263679a0. [DOI] [PubMed] [Google Scholar]
  7. Cortese R., Harland R., Melton D. Transcription of tRNA genes in vivo: single-stranded compared to double-stranded templates. Proc Natl Acad Sci U S A. 1980 Jul;77(7):4147–4151. doi: 10.1073/pnas.77.7.4147. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Denhardt D. T., Kato A. C. Comparison of the effect of ultraviolet radiation and ethidium bromide intercalation on the conformation of superhelical phiX174 replicative form DNA. J Mol Biol. 1973 Jul 15;77(4):479–494. doi: 10.1016/0022-2836(73)90217-9. [DOI] [PubMed] [Google Scholar]
  9. Downes C. S., Ord M. J., Mullinger A. M., Collins A. R., Johnson R. T. Novobiocin inhibition of DNA excision repair may occur through effects on mitochondrial structure and ATP metabolism, not on repair topoisomerases. Carcinogenesis. 1985 Sep;6(9):1343–1352. doi: 10.1093/carcin/6.9.1343. [DOI] [PubMed] [Google Scholar]
  10. Ford C. C., Woodland H. R. DNA synthesis in ocytes and eggs of Xenopus laevis injected with DNA. Dev Biol. 1975 Mar;43(1):189–199. doi: 10.1016/0012-1606(75)90140-2. [DOI] [PubMed] [Google Scholar]
  11. Fuller F. B. The writhing number of a space curve. Proc Natl Acad Sci U S A. 1971 Apr;68(4):815–819. doi: 10.1073/pnas.68.4.815. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Gargiulo G., Worcel A. Analysis of the chromatin assembled in germinal vesicles of Xenopus oocytes. J Mol Biol. 1983 Nov 5;170(3):699–722. doi: 10.1016/s0022-2836(83)80128-4. [DOI] [PubMed] [Google Scholar]
  13. Gurdon J. B., Melton D. A. Gene transfer in amphibian eggs and oocytes. Annu Rev Genet. 1981;15:189–218. doi: 10.1146/annurev.ge.15.120181.001201. [DOI] [PubMed] [Google Scholar]
  14. Gurdon J. B., Wickens M. P. The use of Xenopus oocytes for the expression of cloned genes. Methods Enzymol. 1983;101:370–386. doi: 10.1016/0076-6879(83)01028-9. [DOI] [PubMed] [Google Scholar]
  15. Harland R. M., Weintraub H., McKnight S. L. Transcription of DNA injected into Xenopus oocytes is influenced by template topology. Nature. 1983 Mar 3;302(5903):38–43. doi: 10.1038/302038a0. [DOI] [PubMed] [Google Scholar]
  16. Hsiang Y. H., Hertzberg R., Hecht S., Liu L. F. Camptothecin induces protein-linked DNA breaks via mammalian DNA topoisomerase I. J Biol Chem. 1985 Nov 25;260(27):14873–14878. [PubMed] [Google Scholar]
  17. Kano Y., Fujiwara Y. Defective thymine dimer excision from xeroderma pigmentosum chromatin and its characteristic catalysis by cell-free extracts. Carcinogenesis. 1983 Nov;4(11):1419–1424. doi: 10.1093/carcin/4.11.1419. [DOI] [PubMed] [Google Scholar]
  18. Keyse S. M., Tyrrell R. M. Evidence that novobiocin and nalidixic acid do not inhibit excision repair in u.v.-irradiated human skin fibroblasts at a pre-incision step. Carcinogenesis. 1985 Aug;6(8):1231–1233. doi: 10.1093/carcin/6.8.1231. [DOI] [PubMed] [Google Scholar]
  19. Kressmann A., Clarkson S. G., Pirrotta V., Birnstiel M. L. Transcription of cloned tRNA gene fragments and subfragments injected into the oocyte nucleus of Xenopus laevis. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1176–1180. doi: 10.1073/pnas.75.3.1176. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Legerski R. J., Gray H. B., Jr, Robberson D. L. A sensitive endonuclease probe for lesions in deoxyribonucleic acid helix structure produced by carcinogenic or mutagenic agents. J Biol Chem. 1977 Dec 10;252(23):8740–8746. [PubMed] [Google Scholar]
  21. Mattern M. R., Paone R. F., Day R. S., 3rd Eukaryotic DNA repair is blocked at different steps by inhibitors of DNA topoisomerases and of DNA polymerases alpha and beta. Biochim Biophys Acta. 1982 Apr 26;697(1):6–13. doi: 10.1016/0167-4781(82)90038-0. [DOI] [PubMed] [Google Scholar]
  22. Mattern M. R., Scudiero D. A. Dependence of mammalian DNA synthesis on DNA supercoiling. III. Characterization of the inhibition of replicative and repair-type DNA synthesis by novobiocin and nalidixic acid. Biochim Biophys Acta. 1981 Apr 27;653(2):248–258. doi: 10.1016/0005-2787(81)90160-x. [DOI] [PubMed] [Google Scholar]
  23. Mertz J. E. Linear DNA does not form chromatin containing regularly spaced nucleosomes. Mol Cell Biol. 1982 Dec;2(12):1608–1618. doi: 10.1128/mcb.2.12.1608. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Mortelmans K., Friedberg E. C., Slor H., Thomas G., Cleaver J. E. Defective thymine dimer excision by cell-free extracts of xeroderma pigmentosum cells. Proc Natl Acad Sci U S A. 1976 Aug;73(8):2757–2761. doi: 10.1073/pnas.73.8.2757. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Ryoji M., Worcel A. Chromatin assembly in Xenopus oocytes: in vivo studies. Cell. 1984 May;37(1):21–32. doi: 10.1016/0092-8674(84)90297-6. [DOI] [PubMed] [Google Scholar]
  26. Ryoji M., Worcel A. Structure of the two distinct types of minichromosomes that are assembled on DNA injected in Xenopus oocytes. Cell. 1985 Apr;40(4):923–932. doi: 10.1016/0092-8674(85)90352-6. [DOI] [PubMed] [Google Scholar]
  27. Shure M., Pulleyblank D. E., Vinograd J. The problems of eukaryotic and prokaryotic DNA packaging and in vivo conformation posed by superhelix density heterogeneity. Nucleic Acids Res. 1977;4(5):1183–1205. doi: 10.1093/nar/4.5.1183. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Vinograd J., Lebowitz J. Physical and topological properties of circular DNA. J Gen Physiol. 1966 Jul;49(6):103–125. doi: 10.1085/jgp.49.6.103. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Wang J. C. Helical repeat of DNA in solution. Proc Natl Acad Sci U S A. 1979 Jan;76(1):200–203. doi: 10.1073/pnas.76.1.200. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Wyllie A. H., Laskey R. A., Finch J., Gurdon J. B. Selective DNA conservation and chromatin assembly after injection of SV40 DNA into Xenopus oocytes. Dev Biol. 1978 May;64(1):178–188. doi: 10.1016/0012-1606(78)90069-6. [DOI] [PubMed] [Google Scholar]

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