Skip to main content
PMC 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
. 1979 Jul;76(7):3304–3308. doi: 10.1073/pnas.76.7.3304

Differential sensitivity of gene expression in vitro to inhibitors of DNA gyrase.

H L Yang, K Heller, M Gellert, G Zubay
PMCID: PMC383813  PMID: 226966

Abstract

We have used the antibiotics coumermycin A1, novobiocin, and oxolinic acid, which are specific inhibitors of DNA gyrase, to study the coupled transcription and translation of several bacterial and plasmid genes in a DNA-directed cell-free system. The expression of different genes is reduced to different extents by inhibition of DNA gyrase activity. Among the genes tested, the lac operon, an rRNA gene, and the colicin gene of colicin E1 plasmid were found to be most sensitive, while the trp operon and some other genes in colicin E1 plasmid were relatively unaffected by the inhibitors. These results, together with earlier work on the transcription of circular DNA templates, indicate that DNA supercoiling can significantly enhance transcription from certain promoters.

Full text

PDF
3304

Selected References

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

  1. Barnes W. M. Plasmid detection and sizing in single colony lysates. Science. 1977 Jan 28;195(4276):393–394. doi: 10.1126/science.318764. [DOI] [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. Champoux J. J. Proteins that affect DNA conformation. Annu Rev Biochem. 1978;47:449–479. doi: 10.1146/annurev.bi.47.070178.002313. [DOI] [PubMed] [Google Scholar]
  4. Clewell D. B., Helinski D. R. Supercoiled circular DNA-protein complex in Escherichia coli: purification and induced conversion to an opern circular DNA form. Proc Natl Acad Sci U S A. 1969 Apr;62(4):1159–1166. doi: 10.1073/pnas.62.4.1159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. 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]
  6. Gellert M., Mizuuchi K., O'Dea M. H., Nash H. A. DNA gyrase: an enzyme that introduces superhelical turns into DNA. Proc Natl Acad Sci U S A. 1976 Nov;73(11):3872–3876. doi: 10.1073/pnas.73.11.3872. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. 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]
  8. Hayashi Y., Hayashi M. Template activities of the phi X-174 replicative allomorphic deoxyribonucleic acids. Biochemistry. 1971 Nov;10(23):4212–4218. doi: 10.1021/bi00799a009. [DOI] [PubMed] [Google Scholar]
  9. Herschman H. R., Helinski D. R. Purification and characterization of colicin E2 and colicin E3. J Biol Chem. 1967 Nov 25;242(22):5360–5368. [PubMed] [Google Scholar]
  10. Hershfield V., Boyer H. W., Yanofsky C., Lovett M. A., Helinski D. R. Plasmid ColEl as a molecular vehicle for cloning and amplification of DNA. Proc Natl Acad Sci U S A. 1974 Sep;71(9):3455–3459. doi: 10.1073/pnas.71.9.3455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Itoh T., Tomizawa J. I. Involvement of DNA gyrase in bacteriophage T7 DNA replication. Nature. 1977 Nov 3;270(5632):78–80. doi: 10.1038/270078a0. [DOI] [PubMed] [Google Scholar]
  12. Johnsrud L. Contacts between Escherichia coli RNA polymerase and a lac operon promoter. Proc Natl Acad Sci U S A. 1978 Nov;75(11):5314–5318. doi: 10.1073/pnas.75.11.5314. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kirschbaum J. B., Konrad E. B. Isolation of a specialized lambda transducing bacteriophage carrying the beta subunit gene for Escherichia coli ribonucleic acid polymerase. J Bacteriol. 1973 Nov;116(2):517–526. doi: 10.1128/jb.116.2.517-526.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Lindahl S., Yamamoto M., Nomura M. Mapping of a cluster of genes for components of the transcriptional and translational machineries of Escherichia coli. J Mol Biol. 1977 Jan 5;109(1):23–47. doi: 10.1016/s0022-2836(77)80044-2. [DOI] [PubMed] [Google Scholar]
  15. Puga A., Tessman I. Mechanism of transcription of bacteriophage S13. II. Inhibition of phage-specific transcription by nalidixic acid. J Mol Biol. 1973 Mar 25;75(1):99–108. doi: 10.1016/0022-2836(73)90531-7. [DOI] [PubMed] [Google Scholar]
  16. Richardson J. P. Initiation of transcription by Escherichia coli RNA polymerase from supercoiled and non-supercoiled bacteriophage PM2 DNA. J Mol Biol. 1975 Feb 5;91(4):477–487. doi: 10.1016/0022-2836(75)90274-0. [DOI] [PubMed] [Google Scholar]
  17. Rosner J. L. Formation, induction, and curing of bacteriophage P1 lysogens. Virology. 1972 Jun;48(3):679–689. doi: 10.1016/0042-6822(72)90152-3. [DOI] [PubMed] [Google Scholar]
  18. Saucier J. M., Wang J. C. Angular alteration of the DNA helix by E. coli RNA polymerase. Nat New Biol. 1972 Oct 11;239(93):167–170. doi: 10.1038/newbio239167a0. [DOI] [PubMed] [Google Scholar]
  19. Seeburg P. H., Nüsslein C., Schaller H. Interaction of RNA polymerase with promoters from bacteriophage fd. Eur J Biochem. 1977 Mar 15;74(1):107–113. doi: 10.1111/j.1432-1033.1977.tb11372.x. [DOI] [PubMed] [Google Scholar]
  20. 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]
  21. 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]
  22. Smith D. H., Davis B. D. Mode of action of novobiocin in Escherichia coli. J Bacteriol. 1967 Jan;93(1):71–79. doi: 10.1128/jb.93.1.71-79.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Staudenbauer W. L. Replication of Escherichia coli DNA in vitro: inhibition by oxolinic acid. Eur J Biochem. 1976 Mar 1;62(3):491–497. doi: 10.1111/j.1432-1033.1976.tb10183.x. [DOI] [PubMed] [Google Scholar]
  24. Studier F. W. Analysis of bacteriophage T7 early RNAs and proteins on slab gels. J Mol Biol. 1973 Sep 15;79(2):237–248. doi: 10.1016/0022-2836(73)90003-x. [DOI] [PubMed] [Google Scholar]
  25. 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]
  26. Yang H. L., Zubay G., Levy S. B. Synthesis of an R plasmid protein associated with tetracycline resistance is negatively regulated. Proc Natl Acad Sci U S A. 1976 May;73(5):1509–1512. doi: 10.1073/pnas.73.5.1509. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Yang H., Zubay G. A possible termination factor for transcription in Escherichia coli. Biochem Biophys Res Commun. 1974 Feb 4;56(3):725–731. doi: 10.1016/0006-291x(74)90665-2. [DOI] [PubMed] [Google Scholar]
  28. Zubay G. In vitro synthesis of protein in microbial systems. Annu Rev Genet. 1973;7:267–287. doi: 10.1146/annurev.ge.07.120173.001411. [DOI] [PubMed] [Google Scholar]
  29. Zubay G., Morse D. E., Schrenk W. J., Miller J. H. Detection and isolation of the repressor protein for the tryptophan operon of Escherichia coli. Proc Natl Acad Sci U S A. 1972 May;69(5):1100–1103. doi: 10.1073/pnas.69.5.1100. [DOI] [PMC free article] [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