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. 1981 May;78(5):2747–2751. doi: 10.1073/pnas.78.5.2747

Mutations in the gene coding for Escherichia coli DNA topoisomerase I affect transcription and transposition.

R Sternglanz, S DiNardo, K A Voelkel, Y Nishimura, Y Hirota, K Becherer, L Zumstein, J C Wang
PMCID: PMC319434  PMID: 6265907

Abstract

Mutations in top, the structural gene for Escherichia coli DNA topoisomerase I, have been identified and mapped at 28 min on the chromosome, near cysB. Strains carrying deletions of the top gene are viable. The top mutations, however, do exert pleiotropic effects on transcription and transposition. Mutants lacking DNA topoisomerase I have a more rapid rate of induction and a higher level of catabolite-sensitive enzymes including tryptophanase and beta-galactosidase. This general activation of transcription by top mutations can be attributed to an increase in the negative superhelicity of the DNA in vivo when the topoisomerase activity is abolished. The frequency of transposition of Tn5, a transposon carrying kanamycin resistance, is decreased by a factor of 40 or more in top mutants. A direct or indirect role of the topoisomerase in transposition is discussed. The transposition frequency of Tn3, however, is not dependent on top. Based on the studies of the E. coli top mutants, it appears that the supX gene, which was originally studied in Salmonella typhimurium [Dubnau, E. & Margolin, P. (1972) Mol. Gen. Genet. 117, 91-112] is likely to be the structural gene for DNA topoisomerase I.

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Selected References

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

  1. Bachmann B. J., Low K. B. Linkage map of Escherichia coli K-12, edition 6. Microbiol Rev. 1980 Mar;44(1):1–56. doi: 10.1128/mr.44.1.1-56.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Botchan P. An electron microscopic comparison of transcription on linear and superhelical DNA. J Mol Biol. 1976 Jul 25;105(1):161–176. doi: 10.1016/0022-2836(76)90201-1. [DOI] [PubMed] [Google Scholar]
  3. Botchan P., Wang J. C., Echols H. Effect of circularity and superhelicity on transcription from bacteriophagelambda DNA. Proc Natl Acad Sci U S A. 1973 Nov;70(11):3077–3081. doi: 10.1073/pnas.70.11.3077. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Brown P. O., Peebles C. L., Cozzarelli N. R. A topoisomerase from Escherichia coli related to DNA gyrase. Proc Natl Acad Sci U S A. 1979 Dec;76(12):6110–6114. doi: 10.1073/pnas.76.12.6110. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Clarke L., Carbon J. A colony bank containing synthetic Col El hybrid plasmids representative of the entire E. coli genome. Cell. 1976 Sep;9(1):91–99. doi: 10.1016/0092-8674(76)90055-6. [DOI] [PubMed] [Google Scholar]
  6. Cozzarelli N. R. DNA gyrase and the supercoiling of DNA. Science. 1980 Feb 29;207(4434):953–960. doi: 10.1126/science.6243420. [DOI] [PubMed] [Google Scholar]
  7. Dubnau E., Lenny A. B., Margolin P. Nonsense mutations of the supX locus: further characterization of the supX mutant phenotype. Mol Gen Genet. 1973 Nov 12;126(3):191–200. doi: 10.1007/BF00267530. [DOI] [PubMed] [Google Scholar]
  8. Dubnau E., Margolin P. Suppression of promoter mutations by the pleiotropic supx mutations. Mol Gen Genet. 1972;117(2):91–112. doi: 10.1007/BF00267607. [DOI] [PubMed] [Google Scholar]
  9. Falco S. C., Zivin R., Rothman-Denes L. B. Novel template requirements of N4 virion RNA polymerase. Proc Natl Acad Sci U S A. 1978 Jul;75(7):3220–3224. doi: 10.1073/pnas.75.7.3220. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fimmel A. L., Loughlin R. E. Isolation of a lambdadcys transducing bacteriophage and its use in determining the regulation of cysteine messenger ribonucleic acid synthesis in Escherichia coli K-12. J Bacteriol. 1977 Dec;132(3):757–763. doi: 10.1128/jb.132.3.757-763.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. GARTNER T. K., RILEY M. ISOLATION OF MUTANTS AFFECTING TRYPTOPHANASE PRODUCTION IN ESCHERICHIA COLI. J Bacteriol. 1965 Feb;89:313–318. doi: 10.1128/jb.89.2.313-318.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Gellert M., Mizuuchi K., O'Dea M. H., Itoh T., Tomizawa J. I. Nalidixic acid resistance: a second genetic character involved in DNA gyrase activity. Proc Natl Acad Sci U S A. 1977 Nov;74(11):4772–4776. doi: 10.1073/pnas.74.11.4772. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gellert M., O'Dea M. H., Itoh T., Tomizawa J. Novobiocin and coumermycin inhibit DNA supercoiling catalyzed by DNA gyrase. Proc Natl Acad Sci U S A. 1976 Dec;73(12):4474–4478. doi: 10.1073/pnas.73.12.4474. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Gottesman M. M., Hicks M. L., Gellert M. Genetics and function of DNA ligase in Escherichia coli. J Mol Biol. 1973 Jul 15;77(4):531–547. doi: 10.1016/0022-2836(73)90221-0. [DOI] [PubMed] [Google Scholar]
  15. Heffron F., Bedinger P., Champoux J. J., Falkow S. Deletions affecting the transposition of an antibiotic resistance gene. Proc Natl Acad Sci U S A. 1977 Feb;74(2):702–706. doi: 10.1073/pnas.74.2.702. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Isono S., Isono K. Mutations affecting the structural genes and the genes coding for modifying enzymes for ribosomal proteins in Escherichia coli. Mol Gen Genet. 1978 Sep 20;165(1):15–20. doi: 10.1007/BF00270371. [DOI] [PubMed] [Google Scholar]
  17. Keller W. Characterization of purified DNA-relaxing enzyme from human tissue culture cells. Proc Natl Acad Sci U S A. 1975 Jul;72(7):2550–2554. doi: 10.1073/pnas.72.7.2550. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Keller W., Wendel I. Stepwise relaxation of supercoiled SV40 DNA. Cold Spring Harb Symp Quant Biol. 1975;39(Pt 1):199–208. doi: 10.1101/sqb.1974.039.01.026. [DOI] [PubMed] [Google Scholar]
  19. Kikuchi Y., Nash H. A. Nicking-closing activity associated with bacteriophage lambda int gene product. Proc Natl Acad Sci U S A. 1979 Aug;76(8):3760–3764. doi: 10.1073/pnas.76.8.3760. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kirkegaard K., Wang J. C. Escherichia coli DNA topoisomerase I catalyzed linking of single-stranded rings of complementary base sequences. Nucleic Acids Res. 1978 Oct;5(10):3811–3820. doi: 10.1093/nar/5.10.3811. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Lenny A. B., Margolin P. Locations of the opp and supX genes of Salmonella typhimurium and Escherichia coli. J Bacteriol. 1980 Aug;143(2):747–752. doi: 10.1128/jb.143.2.747-752.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Liu L. F., Depew R. E., Wang J. C. Knotted single-stranded DNA rings: a novel topological isomer of circular single-stranded DNA formed by treatment with Escherichia coli omega protein. J Mol Biol. 1976 Sep 15;106(2):439–452. doi: 10.1016/0022-2836(76)90095-4. [DOI] [PubMed] [Google Scholar]
  23. Mukai F. H., Margolin P. ANALYSIS OF UNLINKED SUPPRESSORS OF AN O degrees MUTATION IN SALMONELLA. Proc Natl Acad Sci U S A. 1963 Jul;50(1):140–148. doi: 10.1073/pnas.50.1.140. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Noguchi S., Yamaizumi Z., Ohgi T., Goto T., Nishimura Y., Hirota Y., Nishimura S. Isolation of Q nucleoside precursor present in tRNA of an E. coli mutant and its characterization as 7-(cyano)-7-deazaguanosine. Nucleic Acids Res. 1978 Nov;5(11):4215–4223. doi: 10.1093/nar/5.11.4215. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Pluschke G., Hirota Y., Overath P. Function of phospholipids in Escherichia coli. Characterization of a mutant deficient in cardiolipin synthesis. J Biol Chem. 1978 Jul 25;253(14):5048–5055. [PubMed] [Google Scholar]
  26. Richardson J. P. Effects of supercoiling on transcription from bacteriophage PM2 deoxyribonucleic acid. Biochemistry. 1974 Jul 16;13(15):3164–3169. doi: 10.1021/bi00712a025. [DOI] [PubMed] [Google Scholar]
  27. Sancar A., Hack A. M., Rupp W. D. Simple method for identification of plasmid-coded proteins. J Bacteriol. 1979 Jan;137(1):692–693. doi: 10.1128/jb.137.1.692-693.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Sankaran L., Pogell B. M. Differential inhibition of catabolite-sensitive enzyme induction by intercalating dyes. Nat New Biol. 1973 Oct 31;245(148):257–260. doi: 10.1038/newbio245257a0. [DOI] [PubMed] [Google Scholar]
  29. Sanzey B. Modulation of gene expression by drugs affecting deoxyribonucleic acid gyrase. J Bacteriol. 1979 Apr;138(1):40–47. doi: 10.1128/jb.138.1.40-47.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Shuman H., Schwartz M. The effect of nalidixic acid on the expression of some genes in Escherichia coli K-12. Biochem Biophys Res Commun. 1975 May 5;64(1):204–209. doi: 10.1016/0006-291x(75)90239-9. [DOI] [PubMed] [Google Scholar]
  31. Smith C. L., Kubo M., Imamoto F. Promoter-specific inhibition of transcription by antibiotics which act on DNA gyrase. Nature. 1978 Oct 5;275(5679):420–423. doi: 10.1038/275420a0. [DOI] [PubMed] [Google Scholar]
  32. Sugino A., Peebles C. L., Kreuzer K. N., Cozzarelli N. R. Mechanism of action of nalidixic acid: purification of Escherichia coli nalA gene product and its relationship to DNA gyrase and a novel nicking-closing enzyme. Proc Natl Acad Sci U S A. 1977 Nov;74(11):4767–4771. doi: 10.1073/pnas.74.11.4767. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Sugiura M., Segawa K., Yoshinaga K., Fujio Y., Ito N. A temperature-sensitive mutant of Escherichia coli with an altered RNA polymerase beta' subunit. Biochem Biophys Res Commun. 1977 Jun 6;76(3):739–745. doi: 10.1016/0006-291x(77)91562-5. [DOI] [PubMed] [Google Scholar]
  34. Sutton A., Freundlich M. Regulation of cyclic AMP of the ilvB-encoded biosynthetic acetohydroxy acid synthase in Escherichia coli K-12. Mol Gen Genet. 1980 Apr;178(1):179–183. doi: 10.1007/BF00267227. [DOI] [PubMed] [Google Scholar]
  35. Suzuki H., Nishimura Y., Hirota Y. On the process of cellular division in Escherichia coli: a series of mutants of E. coli altered in the penicillin-binding proteins. Proc Natl Acad Sci U S A. 1978 Feb;75(2):664–668. doi: 10.1073/pnas.75.2.664. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Tse Y., Wang J. C. E. coli and M. luteus DNA topoisomerase I can catalyze catenation of decatenation of double-stranded DNA rings. Cell. 1980 Nov;22(1 Pt 1):269–276. doi: 10.1016/0092-8674(80)90174-9. [DOI] [PubMed] [Google Scholar]
  37. Wang J. C. Interaction between DNA and an Escherichia coli protein omega. J Mol Biol. 1971 Feb 14;55(3):523–533. doi: 10.1016/0022-2836(71)90334-2. [DOI] [PubMed] [Google Scholar]
  38. Wang J. C. Interactions between twisted DNAs and enzymes: the effects of superhelical turns. J Mol Biol. 1974 Aug 25;87(4):797–816. doi: 10.1016/0022-2836(74)90085-0. [DOI] [PubMed] [Google Scholar]
  39. Yang H. L., Heller K., Gellert M., Zubay G. Differential sensitivity of gene expression in vitro to inhibitors of DNA gyrase. Proc Natl Acad Sci U S A. 1979 Jul;76(7):3304–3308. doi: 10.1073/pnas.76.7.3304. [DOI] [PMC free article] [PubMed] [Google Scholar]

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