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. 1997 Nov 1;25(21):4181–4186. doi: 10.1093/nar/25.21.4181

Processing of topoisomerase I cleavable complexes into DNA damage by transcription.

J Wu 1, L F Liu 1
PMCID: PMC147056  PMID: 9336444

Abstract

Topoisomerase I (TOP1)-mediated DNA damage induced by camptothecin (CPT) in the presence of active transcription has been studied using purified calf thymus TOP1 and T7 RNA polymerase. CPT-stabilized TOP1 cleavable complexes located on the template strand within the transcribed region were found to be converted into irreversible strand breaks by the elongating RNA polymerase. By contrast, CPT-stabilized TOP1 cleavable complexes located on the non-template strand within the transcribed region was unaffected by the elongating RNA polymerase. Previous studies have demonstrated that the elongating T7 RNA polymerase is arrested by TOP1 cleavable complexes located on the template but not the non-template strand [Bendixen et al ., (1990) Biochemistry , 29, 5613-5619]. Together, these results suggest a model in which collision between the TOP1-cleavable complexes located on the template strand and the elongating RNA polymerase results in transcription arrest and conversion of TOP1 cleavable complexes into 'irreversible' strand breaks. The implication of the transcription collision model in DNA damage and repair, as well as cell killing, is discussed.

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Selected References

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  1. Beidler D. R., Cheng Y. C. Camptothecin induction of a time- and concentration-dependent decrease of topoisomerase I and its implication in camptothecin activity. Mol Pharmacol. 1995 May;47(5):907–914. [PubMed] [Google Scholar]
  2. Bendixen C., Thomsen B., Alsner J., Westergaard O. Camptothecin-stabilized topoisomerase I-DNA adducts cause premature termination of transcription. Biochemistry. 1990 Jun 12;29(23):5613–5619. doi: 10.1021/bi00475a028. [DOI] [PubMed] [Google Scholar]
  3. Bonven B. J., Gocke E., Westergaard O. A high affinity topoisomerase I binding sequence is clustered at DNAase I hypersensitive sites in Tetrahymena R-chromatin. Cell. 1985 Jun;41(2):541–551. doi: 10.1016/s0092-8674(85)80027-1. [DOI] [PubMed] [Google Scholar]
  4. Chatterjee S., Cheng M. F., Berger N. A. Hypersensitivity to clinically useful alkylating agents and radiation in poly(ADP-ribose) polymerase-deficient cell lines. Cancer Commun. 1990;2(12):401–407. doi: 10.3727/095535490820873958. [DOI] [PubMed] [Google Scholar]
  5. Chen A. Y., Liu L. F. DNA topoisomerases: essential enzymes and lethal targets. Annu Rev Pharmacol Toxicol. 1994;34:191–218. doi: 10.1146/annurev.pa.34.040194.001203. [DOI] [PubMed] [Google Scholar]
  6. Chen Y. H., Matsumoto Y., Shibutani S., Bogenhagen D. F. Acetylaminofluorene and aminofluorene adducts inhibit in vitro transcription of a Xenopus 5S RNA gene only when located on the coding strand. Proc Natl Acad Sci U S A. 1991 Nov 1;88(21):9583–9587. doi: 10.1073/pnas.88.21.9583. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Christiansen K., Svejstrup A. B., Andersen A. H., Westergaard O. Eukaryotic topoisomerase I-mediated cleavage requires bipartite DNA interaction. Cleavage of DNA substrates containing strand interruptions implicates a role for topoisomerase I in illegitimate recombination. J Biol Chem. 1993 May 5;268(13):9690–9701. [PubMed] [Google Scholar]
  8. D'Arpa P., Beardmore C., Liu L. F. Involvement of nucleic acid synthesis in cell killing mechanisms of topoisomerase poisons. Cancer Res. 1990 Nov 1;50(21):6919–6924. [PubMed] [Google Scholar]
  9. Desai S. D., Liu L. F., Vazquez-Abad D., D'Arpa P. Ubiquitin-dependent destruction of topoisomerase I is stimulated by the antitumor drug camptothecin. J Biol Chem. 1997 Sep 26;272(39):24159–24164. doi: 10.1074/jbc.272.39.24159. [DOI] [PubMed] [Google Scholar]
  10. Donahue B. A., Yin S., Taylor J. S., Reines D., Hanawalt P. C. Transcript cleavage by RNA polymerase II arrested by a cyclobutane pyrimidine dimer in the DNA template. Proc Natl Acad Sci U S A. 1994 Aug 30;91(18):8502–8506. doi: 10.1073/pnas.91.18.8502. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Fleischmann G., Pflugfelder G., Steiner E. K., Javaherian K., Howard G. C., Wang J. C., Elgin S. C. Drosophila DNA topoisomerase I is associated with transcriptionally active regions of the genome. Proc Natl Acad Sci U S A. 1984 Nov;81(22):6958–6962. doi: 10.1073/pnas.81.22.6958. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Froelich-Ammon S. J., Osheroff N. Topoisomerase poisons: harnessing the dark side of enzyme mechanism. J Biol Chem. 1995 Sep 15;270(37):21429–21432. doi: 10.1074/jbc.270.37.21429. [DOI] [PubMed] [Google Scholar]
  13. Gatto B., Sanders M. M., Yu C., Wu H. Y., Makhey D., LaVoie E. J., Liu L. F. Identification of topoisomerase I as the cytotoxic target of the protoberberine alkaloid coralyne. Cancer Res. 1996 Jun 15;56(12):2795–2800. [PubMed] [Google Scholar]
  14. Gilmour D. S., Elgin S. C. Localization of specific topoisomerase I interactions within the transcribed region of active heat shock genes by using the inhibitor camptothecin. Mol Cell Biol. 1987 Jan;7(1):141–148. doi: 10.1128/mcb.7.1.141. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Gilmour D. S., Pflugfelder G., Wang J. C., Lis J. T. Topoisomerase I interacts with transcribed regions in Drosophila cells. Cell. 1986 Feb 14;44(3):401–407. doi: 10.1016/0092-8674(86)90461-7. [DOI] [PubMed] [Google Scholar]
  16. Halligan B. D., Edwards K. A., Liu L. F. Purification and characterization of a type II DNA topoisomerase from bovine calf thymus. J Biol Chem. 1985 Feb 25;260(4):2475–2482. [PubMed] [Google Scholar]
  17. Holm C., Covey J. M., Kerrigan D., Pommier Y. Differential requirement of DNA replication for the cytotoxicity of DNA topoisomerase I and II inhibitors in Chinese hamster DC3F cells. Cancer Res. 1989 Nov 15;49(22):6365–6368. [PubMed] [Google Scholar]
  18. Howard M. T., Neece S. H., Matson S. W., Kreuzer K. N. Disruption of a topoisomerase-DNA cleavage complex by a DNA helicase. Proc Natl Acad Sci U S A. 1994 Dec 6;91(25):12031–12035. doi: 10.1073/pnas.91.25.12031. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. 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]
  20. Hsiang Y. H., Lihou M. G., Liu L. F. Arrest of replication forks by drug-stabilized topoisomerase I-DNA cleavable complexes as a mechanism of cell killing by camptothecin. Cancer Res. 1989 Sep 15;49(18):5077–5082. [PubMed] [Google Scholar]
  21. Hsieh T., Wang J. C. Physiochemical studies on interactions between DNA and RNA polymerase. Ultraviolet absorption measurements. Nucleic Acids Res. 1978 Sep;5(9):3337–3345. doi: 10.1093/nar/5.9.3337. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Huang C. C., Han C. S., Yue X. F., Shen C. M., Wang S. W., Wu F. G., Xu B. Cytotoxicity and sister chromatid exchanges induced in vitro by six anticancer drugs developed in the People's Republic of China. J Natl Cancer Inst. 1983 Oct;71(4):841–847. [PubMed] [Google Scholar]
  23. Kharbanda S., Rubin E., Gunji H., Hinz H., Giovanella B., Pantazis P., Kufe D. Camptothecin and its derivatives induce expression of the c-jun protooncogene in human myeloid leukemia cells. Cancer Res. 1991 Dec 15;51(24):6636–6642. [PubMed] [Google Scholar]
  24. Lim M., Liu L. F., Jacobson-Kram D., Williams J. R. Induction of sister chromatid exchanges by inhibitors of topoisomerases. Cell Biol Toxicol. 1986 Dec;2(4):485–494. doi: 10.1007/BF00117850. [DOI] [PubMed] [Google Scholar]
  25. Liu B., Alberts B. M. Head-on collision between a DNA replication apparatus and RNA polymerase transcription complex. Science. 1995 Feb 24;267(5201):1131–1137. doi: 10.1126/science.7855590. [DOI] [PubMed] [Google Scholar]
  26. Liu L. F. DNA topoisomerase poisons as antitumor drugs. Annu Rev Biochem. 1989;58:351–375. doi: 10.1146/annurev.bi.58.070189.002031. [DOI] [PubMed] [Google Scholar]
  27. Ljungman M., Hanawalt P. C. The anti-cancer drug camptothecin inhibits elongation but stimulates initiation of RNA polymerase II transcription. Carcinogenesis. 1996 Jan;17(1):31–35. doi: 10.1093/carcin/17.1.31. [DOI] [PubMed] [Google Scholar]
  28. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  29. Morris E. J., Geller H. M. Induction of neuronal apoptosis by camptothecin, an inhibitor of DNA topoisomerase-I: evidence for cell cycle-independent toxicity. J Cell Biol. 1996 Aug;134(3):757–770. doi: 10.1083/jcb.134.3.757. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Nelson W. G., Kastan M. B. DNA strand breaks: the DNA template alterations that trigger p53-dependent DNA damage response pathways. Mol Cell Biol. 1994 Mar;14(3):1815–1823. doi: 10.1128/mcb.14.3.1815. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Piret B., Piette J. Topoisomerase poisons activate the transcription factor NF-kappaB in ACH-2 and CEM cells. Nucleic Acids Res. 1996 Nov 1;24(21):4242–4248. doi: 10.1093/nar/24.21.4242. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Porter S. E., Champoux J. J. The basis for camptothecin enhancement of DNA breakage by eukaryotic topoisomerase I. Nucleic Acids Res. 1989 Nov 11;17(21):8521–8532. doi: 10.1093/nar/17.21.8521. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Puvvada M. S., Forrow S. A., Hartley J. A., Stephenson P., Gibson I., Jenkins T. C., Thurston D. E. Inhibition of bacteriophage T7 RNA polymerase in vitro transcription by DNA-binding pyrrolo[2,1-c][1,4]benzodiazepines. Biochemistry. 1997 Mar 4;36(9):2478–2484. doi: 10.1021/bi952490r. [DOI] [PubMed] [Google Scholar]
  34. Ryan A. J., Squires S., Strutt H. L., Evans A., Johnson R. T. Different fates of camptothecin-induced replication fork-associated double-strand DNA breaks in mammalian cells. Carcinogenesis. 1994 May;15(5):823–828. doi: 10.1093/carcin/15.5.823. [DOI] [PubMed] [Google Scholar]
  35. Sastry S. S., Hearst J. E. Studies on the interaction of T7 RNA polymerase with a DNA template containing a site-specifically placed psoralen cross-link. II. Stability and some properties of elongation complexes. J Mol Biol. 1991 Oct 20;221(4):1111–1125. [PubMed] [Google Scholar]
  36. Schultz M. C., Brill S. J., Ju Q., Sternglanz R., Reeder R. H. Topoisomerases and yeast rRNA transcription: negative supercoiling stimulates initiation and topoisomerase activity is required for elongation. Genes Dev. 1992 Jul;6(7):1332–1341. doi: 10.1101/gad.6.7.1332. [DOI] [PubMed] [Google Scholar]
  37. Smith P. J., Makinson T. A., Watson J. V. Enhanced sensitivity to camptothecin in ataxia-telangiectasia cells and its relationship with the expression of DNA topoisomerase I. Int J Radiat Biol. 1989 Feb;55(2):217–231. doi: 10.1080/09553008914550271. [DOI] [PubMed] [Google Scholar]
  38. Squires S., Ryan A. J., Strutt H. L., Johnson R. T. Hypersensitivity of Cockayne's syndrome cells to camptothecin is associated with the generation of abnormally high levels of double strand breaks in nascent DNA. Cancer Res. 1993 May 1;53(9):2012–2019. [PubMed] [Google Scholar]
  39. Stewart A. F., Herrera R. E., Nordheim A. Rapid induction of c-fos transcription reveals quantitative linkage of RNA polymerase II and DNA topoisomerase I enzyme activities. Cell. 1990 Jan 12;60(1):141–149. doi: 10.1016/0092-8674(90)90724-s. [DOI] [PubMed] [Google Scholar]
  40. Tsao Y. P., D'Arpa P., Liu L. F. The involvement of active DNA synthesis in camptothecin-induced G2 arrest: altered regulation of p34cdc2/cyclin B. Cancer Res. 1992 Apr 1;52(7):1823–1829. [PubMed] [Google Scholar]
  41. Tsao Y. P., Russo A., Nyamuswa G., Silber R., Liu L. F. Interaction between replication forks and topoisomerase I-DNA cleavable complexes: studies in a cell-free SV40 DNA replication system. Cancer Res. 1993 Dec 15;53(24):5908–5914. [PubMed] [Google Scholar]
  42. Wang J. C. DNA topoisomerases. Annu Rev Biochem. 1996;65:635–692. doi: 10.1146/annurev.bi.65.070196.003223. [DOI] [PubMed] [Google Scholar]
  43. Wu R. S., Kumar A., Warner J. R. Ribosome formation is blocked by camptothecin, a reversible inhibitor of RNA synthesis. Proc Natl Acad Sci U S A. 1971 Dec;68(12):3009–3014. doi: 10.1073/pnas.68.12.3009. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Zhang H., Wang J. C., Liu L. F. Involvement of DNA topoisomerase I in transcription of human ribosomal RNA genes. Proc Natl Acad Sci U S A. 1988 Feb;85(4):1060–1064. doi: 10.1073/pnas.85.4.1060. [DOI] [PMC free article] [PubMed] [Google Scholar]

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