Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1989 Feb;171(2):674–683. doi: 10.1128/jb.171.2.674-683.1989

Overinitiation of replication of the Escherichia coli chromosome from an integrated runaway-replication derivative of plasmid R1.

R Bernander 1, A Merryweather 1, K Nordström 1
PMCID: PMC209650  PMID: 2644232

Abstract

A 16-base-pair fragment, deletion of which completely inactivated oriC, was replaced by a temperature-dependent runaway-replication derivative (the copy number of which increases with temperature) of the IncFII plasmid R1. The constructed strains were temperature sensitive, and flow cytometry revealed a severalfold increase in the DNA/mass ratio following shifts to nonpermissive temperatures. The cell size distribution was broader in the constructed strains relative to that in the wild type because of asynchrony between the chromosome replication and cell division cycles. This difference was more pronounced for counterclockwise initiation of chromosomal replication, in which small DNA-less cells and long filaments were abundant. Following a temperature shift the cell size distributions became even more broad, showing that changes in the frequency of chromosomal replication affect cell division and emphasizing the interplay between these two processes.

Full text

PDF
674

Images in this article

677Image
on p.677
680Image
on p.680

Selected References

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

  1. Atlung T., Løbner-Olesen A., Hansen F. G. Overproduction of DnaA protein stimulates initiation of chromosome and minichromosome replication in Escherichia coli. Mol Gen Genet. 1987 Jan;206(1):51–59. doi: 10.1007/BF00326535. [DOI] [PubMed] [Google Scholar]
  2. Bird R. E., Louarn J., Martuscelli J., Caro L. Origin and sequence of chromosome replication in Escherichia coli. J Mol Biol. 1972 Oct 14;70(3):549–566. doi: 10.1016/0022-2836(72)90559-1. [DOI] [PubMed] [Google Scholar]
  3. Bolivar F. Construction and characterization of new cloning vehicles. III. Derivatives of plasmid pBR322 carrying unique Eco RI sites for selection of Eco RI generated recombinant DNA molecules. Gene. 1978 Oct;4(2):121–136. doi: 10.1016/0378-1119(78)90025-2. [DOI] [PubMed] [Google Scholar]
  4. Bolivar F., Rodriguez R. L., Greene P. J., Betlach M. C., Heyneker H. L., Boyer H. W., Crosa J. H., Falkow S. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene. 1977;2(2):95–113. [PubMed] [Google Scholar]
  5. Bremer H., Churchward G. Initiation of chromosome replication in Escherichia coli after induction of dnaA gene expression from a lac promoter. J Bacteriol. 1985 Nov;164(2):922–924. doi: 10.1128/jb.164.2.922-924.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Buhk H. J., Messer W. The replication origin region of Escherichia coli: nucleotide sequence and functional units. Gene. 1983 Oct;24(2-3):265–279. doi: 10.1016/0378-1119(83)90087-2. [DOI] [PubMed] [Google Scholar]
  7. Burton P., Holland I. B. Two pathways of division inhibition in UV-irradiated E. coli. Mol Gen Genet. 1983;190(2):309–314. doi: 10.1007/BF00330656. [DOI] [PubMed] [Google Scholar]
  8. Béjar S., Bouché J. P. Molecular cloning of the region of the terminus of Escherichia coli K-12 DNA replication. J Bacteriol. 1983 Feb;153(2):604–609. doi: 10.1128/jb.153.2.604-609.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Cooper S., Helmstetter C. E. Chromosome replication and the division cycle of Escherichia coli B/r. J Mol Biol. 1968 Feb 14;31(3):519–540. doi: 10.1016/0022-2836(68)90425-7. [DOI] [PubMed] [Google Scholar]
  10. Diaz R., Staudenbauer W. L. Origin and direction of mini-R1 plasmid DNA replication in cell extracts of Escherichia coli. J Bacteriol. 1982 Jun;150(3):1077–1084. doi: 10.1128/jb.150.3.1077-1084.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Donachie W. D. Relationship between cell size and time of initiation of DNA replication. Nature. 1968 Sep 7;219(5158):1077–1079. doi: 10.1038/2191077a0. [DOI] [PubMed] [Google Scholar]
  12. Dwek R., Or-Gad S., Rozenhak S., Ron E. Z. Two new cell division mutants in Escherichia coli map near the terminus of chromosome replication. Mol Gen Genet. 1984;193(2):379–381. doi: 10.1007/BF00330697. [DOI] [PubMed] [Google Scholar]
  13. Grinsted J., Saunders J. R., Ingram L. C., Sykes R. B., Richmond M. H. Properties of a R factor which originated in Pseudomonas aeruginosa 1822. J Bacteriol. 1972 May;110(2):529–537. doi: 10.1128/jb.110.2.529-537.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Guerry P., van Embden J., Falkow S. Molecular nature of two nonconjugative plasmids carrying drug resistance genes. J Bacteriol. 1974 Feb;117(2):619–630. doi: 10.1128/jb.117.2.619-630.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Guzman E. C., Jimenez-Sanchez A., Orr E., Pritchard R. H. Heat stress in the presence of low RNA polymerase activity increases chromosome copy number of Escherichia coli. Mol Gen Genet. 1988 May;212(2):203–206. doi: 10.1007/BF00334685. [DOI] [PubMed] [Google Scholar]
  16. Helmstetter C. E., Krajewski C. A., Leonard A. C., Weinberger M. Discontinuity in DNA replication during expression of accumulated initiation potential in dnaA mutants of Escherichia coli. J Bacteriol. 1986 Feb;165(2):631–637. doi: 10.1128/jb.165.2.631-637.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Helmstetter C. E., Pierucci O. Cell division during inhibition of deoxyribonucleic acid synthesis in Escherichia coli. J Bacteriol. 1968 May;95(5):1627–1633. doi: 10.1128/jb.95.5.1627-1633.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hussain K., Begg K. J., Salmond G. P., Donachie W. D. ParD: a new gene coding for a protein required for chromosome partitioning and septum localization in Escherichia coli. Mol Microbiol. 1987 Jul;1(1):73–81. doi: 10.1111/j.1365-2958.1987.tb00529.x. [DOI] [PubMed] [Google Scholar]
  19. Hussain K., Elliott E. J., Salmond G. P. The parD- mutant of Escherichia coli also carries a gyrAam mutation. The complete sequence of gyrA. Mol Microbiol. 1987 Nov;1(3):259–273. doi: 10.1111/j.1365-2958.1987.tb01932.x. [DOI] [PubMed] [Google Scholar]
  20. Kellenberger-Gujer G., Podhajska A. J., Caro L. A cold sensitive dnaA mutant of E. coli which overinitiates chromosome replication at low temperature. Mol Gen Genet. 1978 Jun 1;162(1):9–16. doi: 10.1007/BF00333845. [DOI] [PubMed] [Google Scholar]
  21. Koppes L., Nordström K. Insertion of an R1 plasmid into the origin of replication of the E. coli chromosome: random timing of replication of the hybrid chromosome. Cell. 1986 Jan 17;44(1):117–124. doi: 10.1016/0092-8674(86)90490-3. [DOI] [PubMed] [Google Scholar]
  22. Larsen J. E., Gerdes K., Light J., Molin S. Low-copy-number plasmid-cloning vectors amplifiable by derepression of an inserted foreign promoter. Gene. 1984 Apr;28(1):45–54. doi: 10.1016/0378-1119(84)90086-6. [DOI] [PubMed] [Google Scholar]
  23. Masai H., Kaziro Y., Arai K. Definition of oriR, the minimum DNA segment essential for initiation of R1 plasmid replication in vitro. Proc Natl Acad Sci U S A. 1983 Nov;80(22):6814–6818. doi: 10.1073/pnas.80.22.6814. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Merryweather A., Bernander R., Nordström K. Direct selection for the exchange of alleles between a plasmid and the Escherichia coli chromosome. Mol Gen Genet. 1987 Dec;210(3):381–384. doi: 10.1007/BF00327186. [DOI] [PubMed] [Google Scholar]
  25. Molin S., Diaz R., Uhlin B. E., Nordström K. Runaway replication of plasmid R1 is not caused by loss of replication inhibitor activity of gene cop. J Bacteriol. 1980 Aug;143(2):1046–1048. doi: 10.1128/jb.143.2.1046-1048.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Nordström K. Control of plasmid replication: theoretical considerations and practical solutions. Basic Life Sci. 1985;30:189–214. doi: 10.1007/978-1-4613-2447-8_17. [DOI] [PubMed] [Google Scholar]
  27. Pierucci O., Helmstetter C. E., Rickert M., Weinberger M., Leonard A. C. Overexpression of the dnaA gene in Escherichia coli B/r: chromosome and minichromosome replication in the presence of rifampin. J Bacteriol. 1987 May;169(5):1871–1877. doi: 10.1128/jb.169.5.1871-1877.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Prentki P., Krisch H. M. In vitro insertional mutagenesis with a selectable DNA fragment. Gene. 1984 Sep;29(3):303–313. doi: 10.1016/0378-1119(84)90059-3. [DOI] [PubMed] [Google Scholar]
  29. Prescott D. M., Kuempel P. L. Bidirectional replication of the chromosome in Escherichia coli. Proc Natl Acad Sci U S A. 1972 Oct;69(10):2842–2845. doi: 10.1073/pnas.69.10.2842. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Rasmussen K. V., Atlung T., Kerszman G., Hansen G. E., Hansen F. G. Conditional change of DNA replication control in an RNA polymerase mutant of Escherichia coli. J Bacteriol. 1983 Apr;154(1):443–451. doi: 10.1128/jb.154.1.443-451.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Ruusala T., Andersson D., Ehrenberg M., Kurland C. G. Hyper-accurate ribosomes inhibit growth. EMBO J. 1984 Nov;3(11):2575–2580. doi: 10.1002/j.1460-2075.1984.tb02176.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Scott J. R. Regulation of plasmid replication. Microbiol Rev. 1984 Mar;48(1):1–23. doi: 10.1016/b978-0-12-048850-6.50006-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Skarstad K., Boye E. Perturbed chromosomal replication in recA mutants of Escherichia coli. J Bacteriol. 1988 Jun;170(6):2549–2554. doi: 10.1128/jb.170.6.2549-2554.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Skarstad K., Boye E., Steen H. B. Timing of initiation of chromosome replication in individual Escherichia coli cells. EMBO J. 1986 Jul;5(7):1711–1717. doi: 10.1002/j.1460-2075.1986.tb04415.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Skarstad K., von Meyenburg K., Hansen F. G., Boye E. Coordination of chromosome replication initiation in Escherichia coli: effects of different dnaA alleles. J Bacteriol. 1988 Feb;170(2):852–858. doi: 10.1128/jb.170.2.852-858.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Tanaka M., Ohmori H., Hiraga S. A novel type of E. coli mutants with increased chromosomal copy number. Mol Gen Genet. 1983;192(1-2):51–60. doi: 10.1007/BF00327646. [DOI] [PubMed] [Google Scholar]
  37. Tormo A., Dopazo A., de la Campa A. G., Aldea M., Vicente M. Coupling between DNA replication and cell division mediated by the FtsA protein in Escherichia coli: a pathway independent of the SOS response, the "TER" pathway. J Bacteriol. 1985 Nov;164(2):950–953. doi: 10.1128/jb.164.2.950-953.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Tormo A., Fernández-Cabrera C., Vicente M. The ftsA gene product: a possible connection between DNA replication and septation in Escherichia coli. J Gen Microbiol. 1985 Feb;131(2):239–244. doi: 10.1099/00221287-131-2-239. [DOI] [PubMed] [Google Scholar]
  39. Xu Y. C., Bremer H. Chromosome replication in Escherichia coli induced by oversupply of DnaA. Mol Gen Genet. 1988 Jan;211(1):138–142. doi: 10.1007/BF00338404. [DOI] [PubMed] [Google Scholar]
  40. von Meyenburg K., Boye E., Skarstad K., Koppes L., Kogoma T. Mode of initiation of constitutive stable DNA replication in RNase H-defective mutants of Escherichia coli K-12. J Bacteriol. 1987 Jun;169(6):2650–2658. doi: 10.1128/jb.169.6.2650-2658.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES