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J Bacteriol. Jun 1997; 179(11): 3528–3533.
PMCID: PMC179144

In vivo supercoiling of plasmid and chromosomal DNA in an Escherichia coli hns mutant.


We have used trimethylpsoralen to measure localized levels of unconstrained DNA supercoiling in vivo. The data provide direct evidence that plasmid and chromosomal DNA supercoiling is altered in vivo in an hns mutant. This increase in supercoiling is independent of transcription or changes in the activity of topoisomerase I. These data have implications for the mechanisms by which the chromatin-associated protein H-NS may influence chromosome organization and gene expression.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Bliska JB, Cozzarelli NR. Use of site-specific recombination as a probe of DNA structure and metabolism in vivo. J Mol Biol. 1987 Mar 20;194(2):205–218. [PubMed]
  • Chen D, Bowater R, Dorman CJ, Lilley DM. Activity of a plasmid-borne leu-500 promoter depends on the transcription and translation of an adjacent gene. Proc Natl Acad Sci U S A. 1992 Sep 15;89(18):8784–8788. [PMC free article] [PubMed]
  • Chen D, Bowater R, Lilley DM. Topological promoter coupling in Escherichia coli: delta topA-dependent activation of the leu-500 promoter on a plasmid. J Bacteriol. 1994 Jun;176(12):3757–3764. [PMC free article] [PubMed]
  • Cook DN, Armstrong GA, Hearst JE. Induction of anaerobic gene expression in Rhodobacter capsulatus is not accompanied by a local change in chromosomal supercoiling as measured by a novel assay. J Bacteriol. 1989 Sep;171(9):4836–4843. [PMC free article] [PubMed]
  • Dri AM, Moreau PL, Rouvière-Yaniv J. Role of the histone-like proteins OsmZ and HU in homologous recombination. Gene. 1992 Oct 12;120(1):11–16. [PubMed]
  • Drlica K. Control of bacterial DNA supercoiling. Mol Microbiol. 1992 Feb;6(4):425–433. [PubMed]
  • Drlica K, Rouviere-Yaniv J. Histonelike proteins of bacteria. Microbiol Rev. 1987 Sep;51(3):301–319. [PMC free article] [PubMed]
  • Drlica K, Franco RJ, Steck TR. Rifampin and rpoB mutations can alter DNA supercoiling in Escherichia coli. J Bacteriol. 1988 Oct;170(10):4983–4985. [PMC free article] [PubMed]
  • Göransson M, Sondén B, Nilsson P, Dagberg B, Forsman K, Emanuelsson K, Uhlin BE. Transcriptional silencing and thermoregulation of gene expression in Escherichia coli. Nature. 1990 Apr 12;344(6267):682–685. [PubMed]
  • Higgins CF, Dorman CJ, Stirling DA, Waddell L, Booth IR, May G, Bremer E. A physiological role for DNA supercoiling in the osmotic regulation of gene expression in S. typhimurium and E. coli. Cell. 1988 Feb 26;52(4):569–584. [PubMed]
  • Higgins CF, Hinton JC, Hulton CS, Owen-Hughes T, Pavitt GD, Seirafi A. Protein H1: a role for chromatin structure in the regulation of bacterial gene expression and virulence? Mol Microbiol. 1990 Dec;4(12):2007–2012. [PubMed]
  • Hinton JC, Santos DS, Seirafi A, Hulton CS, Pavitt GD, Higgins CF. Expression and mutational analysis of the nucleoid-associated protein H-NS of Salmonella typhimurium. Mol Microbiol. 1992 Aug;6(16):2327–2337. [PubMed]
  • Hulton CS, Seirafi A, Hinton JC, Sidebotham JM, Waddell L, Pavitt GD, Owen-Hughes T, Spassky A, Buc H, Higgins CF. Histone-like protein H1 (H-NS), DNA supercoiling, and gene expression in bacteria. Cell. 1990 Nov 2;63(3):631–642. [PubMed]
  • Jordi BJ, Owen-Hughes TA, Hulton CS, Higgins CF. DNA twist, flexibility and transcription of the osmoregulated proU promoter of Salmonella typhimurium. EMBO J. 1995 Nov 15;14(22):5690–5700. [PMC free article] [PubMed]
  • Lejeune P, Danchin A. Mutations in the bglY gene increase the frequency of spontaneous deletions in Escherichia coli K-12. Proc Natl Acad Sci U S A. 1990 Jan;87(1):360–363. [PMC free article] [PubMed]
  • Lilley D. Bacterial chromatin. A new twist to an old story. Nature. 1986 Mar 6;320(6057):14–15. [PubMed]
  • Liu LF, Wang JC. Supercoiling of the DNA template during transcription. Proc Natl Acad Sci U S A. 1987 Oct;84(20):7024–7027. [PMC free article] [PubMed]
  • Lodge JK, Kazic T, Berg DE. Formation of supercoiling domains in plasmid pBR322. J Bacteriol. 1989 Apr;171(4):2181–2187. [PMC free article] [PubMed]
  • Lynch AS, Wang JC. Anchoring of DNA to the bacterial cytoplasmic membrane through cotranscriptional synthesis of polypeptides encoding membrane proteins or proteins for export: a mechanism of plasmid hypernegative supercoiling in mutants deficient in DNA topoisomerase I. J Bacteriol. 1993 Mar;175(6):1645–1655. [PMC free article] [PubMed]
  • May G, Dersch P, Haardt M, Middendorf A, Bremer E. The osmZ (bglY) gene encodes the DNA-binding protein H-NS (H1a), a component of the Escherichia coli K12 nucleoid. Mol Gen Genet. 1990 Oct;224(1):81–90. [PubMed]
  • McLaren RS, Newbury SF, Dance GS, Causton HC, Higgins CF. mRNA degradation by processive 3'-5' exoribonucleases in vitro and the implications for prokaryotic mRNA decay in vivo. J Mol Biol. 1991 Sep 5;221(1):81–95. [PubMed]
  • Mojica FJ, Charbonnier F, Juez G, Rodríguez-Valera F, Forterre P. Effects of salt and temperature on plasmid topology in the halophilic archaeon Haloferax volcanii. J Bacteriol. 1994 Aug;176(16):4966–4973. [PMC free article] [PubMed]
  • Mojica FJ, Higgins CF. Localized domains of DNA supercoiling: topological coupling between promoters. Mol Microbiol. 1996 Dec;22(5):919–928. [PubMed]
  • Ni Bhriain N, Dorman CJ, Higgins CF. An overlap between osmotic and anaerobic stress responses: a potential role for DNA supercoiling in the coordinate regulation of gene expression. Mol Microbiol. 1989 Jul;3(7):933–942. [PubMed]
  • Owen-Hughes TA, Pavitt GD, Santos DS, Sidebotham JM, Hulton CS, Hinton JC, Higgins CF. The chromatin-associated protein H-NS interacts with curved DNA to influence DNA topology and gene expression. Cell. 1992 Oct 16;71(2):255–265. [PubMed]
  • Pavitt GD, Higgins CF. Chromosomal domains of supercoiling in Salmonella typhimurium. Mol Microbiol. 1993 Nov;10(3):685–696. [PubMed]
  • Pruss GJ, Drlica K. DNA supercoiling and suppression of the leu-500 promoter mutation. J Bacteriol. 1985 Nov;164(2):947–949. [PMC free article] [PubMed]
  • Pruss GJ, Manes SH, Drlica K. Escherichia coli DNA topoisomerase I mutants: increased supercoiling is corrected by mutations near gyrase genes. Cell. 1982 Nov;31(1):35–42. [PubMed]
  • Pruss GJ, Drlica K. Topoisomerase I mutants: the gene on pBR322 that encodes resistance to tetracycline affects plasmid DNA supercoiling. Proc Natl Acad Sci U S A. 1986 Dec;83(23):8952–8956. [PMC free article] [PubMed]
  • Py B, Higgins CF, Krisch HM, Carpousis AJ. A DEAD-box RNA helicase in the Escherichia coli RNA degradosome. Nature. 1996 May 9;381(6578):169–172. [PubMed]
  • Rahmouni AR, Wells RD. Stabilization of Z DNA in vivo by localized supercoiling. Science. 1989 Oct 20;246(4928):358–363. [PubMed]
  • Raji A, Zabel DJ, Laufer CS, Depew RE. Genetic analysis of mutations that compensate for loss of Escherichia coli DNA topoisomerase I. J Bacteriol. 1985 Jun;162(3):1173–1179. [PMC free article] [PubMed]
  • Reed KC, Mann DA. Rapid transfer of DNA from agarose gels to nylon membranes. Nucleic Acids Res. 1985 Oct 25;13(20):7207–7221. [PMC free article] [PubMed]
  • Richardson SM, Higgins CF, Lilley DM. DNA supercoiling and the leu-500 promoter mutation of Salmonella typhimurium. EMBO J. 1988 Jun;7(6):1863–1869. [PMC free article] [PubMed]
  • Sinden RR, Carlson JO, Pettijohn DE. Torsional tension in the DNA double helix measured with trimethylpsoralen in living E. coli cells: analogous measurements in insect and human cells. Cell. 1980 Oct;21(3):773–783. [PubMed]
  • Sinden RR, Pettijohn DE. Chromosomes in living Escherichia coli cells are segregated into domains of supercoiling. Proc Natl Acad Sci U S A. 1981 Jan;78(1):224–228. [PMC free article] [PubMed]
  • Sinden RR, Ussery DW. Analysis of DNA structure in vivo using psoralen photobinding: measurement of supercoiling, topological domains, and DNA-protein interactions. Methods Enzymol. 1992;212:319–335. [PubMed]
  • Stirling DA, Hulton CS, Waddell L, Park SF, Stewart GS, Booth IR, Higgins CF. Molecular characterization of the proU loci of Salmonella typhimurium and Escherichia coli encoding osmoregulated glycine betaine transport systems. Mol Microbiol. 1989 Aug;3(8):1025–1038. [PubMed]
  • Tupper AE, Owen-Hughes TA, Ussery DW, Santos DS, Ferguson DJ, Sidebotham JM, Hinton JC, Higgins CF. The chromatin-associated protein H-NS alters DNA topology in vitro. EMBO J. 1994 Jan 1;13(1):258–268. [PMC free article] [PubMed]
  • Worcel A, Burgi E. On the structure of the folded chromosome of Escherichia coli. J Mol Biol. 1972 Nov 14;71(2):127–147. [PubMed]
  • Yamada H, Yoshida T, Tanaka K, Sasakawa C, Mizuno T. Molecular analysis of the Escherichia coli hns gene encoding a DNA-binding protein, which preferentially recognizes curved DNA sequences. Mol Gen Genet. 1991 Nov;230(1-2):332–336. [PubMed]
  • Zheng GX, Kochel T, Hoepfner RW, Timmons SE, Sinden RR. Torsionally tuned cruciform and Z-DNA probes for measuring unrestrained supercoiling at specific sites in DNA of living cells. J Mol Biol. 1991 Sep 5;221(1):107–122. [PubMed]

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