Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1979 Feb;137(2):885–890. doi: 10.1128/jb.137.2.885-890.1979

Biochemical characterization of nonintegrated plasmid-folded chromosome complexes: sex factor F and the Escherichia coli nucleoid.

J R Miller, B C Kline
PMCID: PMC218371  PMID: 370105

Abstract

The existence of nonintegrated plasmid-chromosome complexes has been deduced in previous work from the cosedimentation of covalently closed, circular plasmids with host folded chromosomes. In the present work, it is shown that about 70 to 90% of the covalently closed, circular F deoxyribonucleic acid could be released in vitro from chromosome complexes by ribonuclease treatment but not by protease, Sarkosyl, or ethidium bromide. Consistent with the in vitro studies, Escherichia coli cells treated for 5 min with rifampin, an inhibitor of ribonucleic acid initiation, released upon lysis 90% of their plasmid deoxyribonucleic acid as freely sedimenting molecules.

Full text

PDF
885

Selected References

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

  1. Archibold E. R., Clark C. W., Sheehy R. J. Relationship of R6K replicating forms to the folded chromosome of Escherichia coli. J Bacteriol. 1978 Aug;135(2):476–482. doi: 10.1128/jb.135.2.476-482.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Drlica K., Worcel A. Conformational transitions in the Escherichia coli chromosome: analysis by viscometry and sedimentation. J Mol Biol. 1975 Oct 25;98(2):393–411. doi: 10.1016/s0022-2836(75)80126-4. [DOI] [PubMed] [Google Scholar]
  3. Hansen J. B., Olsen R. H. Isolation of large bacterial plasmids and characterization of the P2 incompatibility group plasmids pMG1 and pMG5. J Bacteriol. 1978 Jul;135(1):227–238. doi: 10.1128/jb.135.1.227-238.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Hecht R. M., Pettijohn D. E. Studies of DNA bound RNA molecules isolated from nucleoids of Escherichia coli. Nucleic Acids Res. 1976 Mar;3(3):767–788. doi: 10.1093/nar/3.3.767. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Hecht R. M., Stimpson D., Pettijohn D. Sedimentation properties of the bacterial chromosome as an isolated nucleoid and as an unfolded DNA fiber. Chromosomal DNA folding measured by rotor speed effects. J Mol Biol. 1977 Apr 15;111(3):257–277. doi: 10.1016/s0022-2836(77)80051-x. [DOI] [PubMed] [Google Scholar]
  6. Hershfield V., LeBlanc D. J., Falkow S. Membrane attachment of R-factor deoxyribonucleic acid in compatible and incompatible cell pairs following conjugation. J Bacteriol. 1973 Sep;115(3):1208–1211. doi: 10.1128/jb.115.3.1208-1211.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Kavenoff R., Bowen B. C. Electron microscopy of membrane-free folded chromosomes from Escherichia coli. Chromosoma. 1976 Dec 16;59(2):89–101. doi: 10.1007/BF00328479. [DOI] [PubMed] [Google Scholar]
  8. Kline B. C., Helinski D. R. F 1 sex factor of Escherichia coli. Size and purification in the form of a strand-specific relaxation complex of supercoiled deoxyribonucleic acid and protein. Biochemistry. 1971 Dec 21;10(26):4975–4980. doi: 10.1021/bi00802a022. [DOI] [PubMed] [Google Scholar]
  9. Kline B. C., Miller J. R., Cress D. E., Wlodarczyk M., Manis J. J., Otten M. R. Nonintegrated plasmid-chromosome complexes in Escherichia coli. J Bacteriol. 1976 Aug;127(2):881–889. doi: 10.1128/jb.127.2.881-889.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kline B. C., Miller J. R. Detection of nonintegrated plasmid deoxyribonucleic acid in the folded chromosome of Escherichia coli: physiochemical approach to studying the unit of segregation. J Bacteriol. 1975 Jan;121(1):165–172. doi: 10.1128/jb.121.1.165-172.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Miller J., Manis J., Kline B., Bishop A. Nonintegrated plasmid-folded chromosome complexes: genetic studies on formation and possible relationship to plasmid replication. Plasmid. 1978 Jun;1(3):273–283. doi: 10.1016/0147-619x(78)90045-8. [DOI] [PubMed] [Google Scholar]
  12. Palchaudhuri S., Chakrabarty A. Isolation of plasmid deoxyribonucleic acid from Pseudomonas putida. J Bacteriol. 1976 Apr;126(1):410–416. doi: 10.1128/jb.126.1.410-416.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Pettijohn D. E., Hecht R. RNA molecules bound to the folded bacterial genome stabilize DNA folds and segregate domains of supercoiling. Cold Spring Harb Symp Quant Biol. 1974;38:31–41. doi: 10.1101/sqb.1974.038.01.006. [DOI] [PubMed] [Google Scholar]
  14. Pritchard R. H., Chandler M. G., Collins J. Independence of F replication and chromosome replication in Escherichia coli. Mol Gen Genet. 1975;138(2):143–155. doi: 10.1007/BF02428118. [DOI] [PubMed] [Google Scholar]
  15. Radloff R., Bauer W., Vinograd J. A dye-buoyant-density method for the detection and isolation of closed circular duplex DNA: the closed circular DNA in HeLa cells. Proc Natl Acad Sci U S A. 1967 May;57(5):1514–1521. doi: 10.1073/pnas.57.5.1514. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Stonington O. G., Pettijohn D. E. The folded genome of Escherichia coli isolated in a protein-DNA-RNA complex. Proc Natl Acad Sci U S A. 1971 Jan;68(1):6–9. doi: 10.1073/pnas.68.1.6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Taichman L., Rownd R. H. Differential association of F' plasmid and R plasmid deoxyribonucleic acid with a rapidly sedimenting fraction of a Proteus mirabilis lysate. J Bacteriol. 1977 Jun;130(3):1262–1273. doi: 10.1128/jb.130.3.1262-1273.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Wlodarczyk M., Kline B. C. Association of R6K plasmid replicative intermediates with the folded chromosome of Escherichia coli. Biochem Biophys Res Commun. 1976 Nov 22;73(2):286–292. doi: 10.1016/0006-291x(76)90705-1. [DOI] [PubMed] [Google Scholar]
  19. Worcel A., Burgi E. On the structure of the folded chromosome of Escherichia coli. J Mol Biol. 1972 Nov 14;71(2):127–147. doi: 10.1016/0022-2836(72)90342-7. [DOI] [PubMed] [Google Scholar]
  20. Worcel A., Burgi E. Properties of a membrane-attached form of the folded chromosome of Escherichia coli. J Mol Biol. 1974 Jan 5;82(1):91–105. doi: 10.1016/0022-2836(74)90576-2. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES