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. 1995 Apr;177(7):1670–1682. doi: 10.1128/jb.177.7.1670-1682.1995

Inhibition and restart of initiation of chromosome replication: effects on exponentially growing Escherichia coli cells.

R Bernander 1, T Akerlund 1, K Nordström 1
PMCID: PMC176792  PMID: 7896687

Abstract

Escherichia coli strains in which initiation of chromosome replication could be specifically blocked while other cellular processes continued uninhibited were constructed. Inhibition of replication resulted in a reduced growth rate and in inhibition of cell division after a time period roughly corresponding to the sum of the lengths of the C and D periods. The division inhibition was not mediated by the SOS regulon. The cells became elongated, and a majority contained a centrally located nucleoid with a fully replicated chromosome. The replication block was reversible, and restart of chromosome replication allowed cell division and rapid growth to resume after a time delay. After the resumption, the septum positions were nonrandomly distributed along the length axis of the cells, and a majority of the divisions resulted in at least one newborn cell of normal size and DNA content. With a transient temperature shift, a single synchronous round of chromosome replication and cell division could be induced in the population, making the constructed system useful for studies of cell cycle-specific events. The coordination between chromosome replication, nucleoid segregation, and cell division in E. coli is discussed.

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Selected References

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  1. Akerlund T., Bernander R., Nordström K. Cell division in Escherichia coli minB mutants. Mol Microbiol. 1992 Aug;6(15):2073–2083. doi: 10.1111/j.1365-2958.1992.tb01380.x. [DOI] [PubMed] [Google Scholar]
  2. Akerlund T., Nordström K., Bernander R. Branched Escherichia coli cells. Mol Microbiol. 1993 Nov;10(4):849–858. doi: 10.1111/j.1365-2958.1993.tb00955.x. [DOI] [PubMed] [Google Scholar]
  3. Allman R., Schjerven T., Boye E. Cell cycle parameters of Escherichia coli K-12. J Bacteriol. 1991 Dec;173(24):7970–7974. doi: 10.1128/jb.173.24.7970-7974.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Begg K. J., Donachie W. D. Experiments on chromosome separation and positioning in Escherichia coli. New Biol. 1991 May;3(5):475–486. [PubMed] [Google Scholar]
  5. Bernander R., Merryweather A., Nordström K. Overinitiation of replication of the Escherichia coli chromosome from an integrated runaway-replication derivative of plasmid R1. J Bacteriol. 1989 Feb;171(2):674–683. doi: 10.1128/jb.171.2.674-683.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bernander R. Universal cell cycle regulation? Trends Cell Biol. 1994 Mar;4(3):76–79. doi: 10.1016/0962-8924(94)90173-2. [DOI] [PubMed] [Google Scholar]
  7. Bi E., Lutkenhaus J. Cell division inhibitors SulA and MinCD prevent formation of the FtsZ ring. J Bacteriol. 1993 Feb;175(4):1118–1125. doi: 10.1128/jb.175.4.1118-1125.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. 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]
  9. Campbell J. L., Kleckner N. E. coli oriC and the dnaA gene promoter are sequestered from dam methyltransferase following the passage of the chromosomal replication fork. Cell. 1990 Sep 7;62(5):967–979. doi: 10.1016/0092-8674(90)90271-f. [DOI] [PubMed] [Google Scholar]
  10. D'Ari R., Huisman O. Novel mechanism of cell division inhibition associated with the SOS response in Escherichia coli. J Bacteriol. 1983 Oct;156(1):243–250. doi: 10.1128/jb.156.1.243-250.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Dam M., Gerdes K. Partitioning of plasmid R1. Ten direct repeats flanking the parA promoter constitute a centromere-like partition site parC, that expresses incompatibility. J Mol Biol. 1994 Mar 11;236(5):1289–1298. doi: 10.1016/0022-2836(94)90058-2. [DOI] [PubMed] [Google Scholar]
  12. Diaz R., Nordström K., Staudenbauer W. L. Plasmid R1 DNA replication dependent on protein synthesis in cell-free extracts of E. coli. Nature. 1981 Jan 22;289(5795):326–328. doi: 10.1038/289326a0. [DOI] [PubMed] [Google Scholar]
  13. Donachie W. D., Begg K. J. Cell length, nucleoid separation, and cell division of rod-shaped and spherical cells of Escherichia coli. J Bacteriol. 1989 Sep;171(9):4633–4639. doi: 10.1128/jb.171.9.4633-4639.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Donachie W. D. Control of cell division in Escherichia coli: experiments with thymine starvation. J Bacteriol. 1969 Oct;100(1):260–268. doi: 10.1128/jb.100.1.260-268.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Donachie W. D., Hobbs D. G., Masters M. Chromosome replication and cell division in Escherichia coli 15T after growth in the absence of DNA synthesis. Nature. 1968 Sep 7;219(5158):1079–1080. doi: 10.1038/2191079a0. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. Gustafsson P., Nordström K., Perram J. W. Selection and timing of replication of plasmids R1drd-19 and F'lac in Escherichia coli. Plasmid. 1978 Feb;1(2):187–203. doi: 10.1016/0147-619x(78)90038-0. [DOI] [PubMed] [Google Scholar]
  18. Hansen F. G., Christensen B. B., Atlung T. The initiator titration model: computer simulation of chromosome and minichromosome control. Res Microbiol. 1991 Feb-Apr;142(2-3):161–167. doi: 10.1016/0923-2508(91)90025-6. [DOI] [PubMed] [Google Scholar]
  19. Heitman J., Model P. SOS induction as an in vivo assay of enzyme-DNA interactions. Gene. 1991 Jul 15;103(1):1–9. doi: 10.1016/0378-1119(91)90383-m. [DOI] [PubMed] [Google Scholar]
  20. Helmstetter C. E., Pierucci O. DNA synthesis during the division cycle of three substrains of Escherichia coli B/r. J Mol Biol. 1976 Apr 15;102(3):477–486. doi: 10.1016/0022-2836(76)90329-6. [DOI] [PubMed] [Google Scholar]
  21. Helmstetter C., Cooper S., Pierucci O., Revelas E. On the bacterial life sequence. Cold Spring Harb Symp Quant Biol. 1968;33:809–822. doi: 10.1101/sqb.1968.033.01.093. [DOI] [PubMed] [Google Scholar]
  22. Hiraga S., Ogura T., Niki H., Ichinose C., Mori H. Positioning of replicated chromosomes in Escherichia coli. J Bacteriol. 1990 Jan;172(1):31–39. doi: 10.1128/jb.172.1.31-39.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Hirota Y., Jacob F., Ryter A., Buttin G., Nakai T. On the process of cellular division in Escherichia coli. I. Asymmetrical cell division and production of deoxyribonucleic acid-less bacteria. J Mol Biol. 1968 Jul 14;35(1):175–192. doi: 10.1016/s0022-2836(68)80046-4. [DOI] [PubMed] [Google Scholar]
  24. Huisman O., D'Ari R. An inducible DNA replication-cell division coupling mechanism in E. coli. Nature. 1981 Apr 30;290(5809):797–799. doi: 10.1038/290797a0. [DOI] [PubMed] [Google Scholar]
  25. Jaffé A., D'Ari R., Norris V. SOS-independent coupling between DNA replication and cell division in Escherichia coli. J Bacteriol. 1986 Jan;165(1):66–71. doi: 10.1128/jb.165.1.66-71.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Jensen K. F. The Escherichia coli K-12 "wild types" W3110 and MG1655 have an rph frameshift mutation that leads to pyrimidine starvation due to low pyrE expression levels. J Bacteriol. 1993 Jun;175(11):3401–3407. doi: 10.1128/jb.175.11.3401-3407.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Jones N. C., Donachie W. D. Protein synthesis and the release of the replicated chromosome from the cell membrane. Nature. 1974 Sep 20;251(5472):252–254. doi: 10.1038/251252a0. [DOI] [PubMed] [Google Scholar]
  28. 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]
  29. LARK K. G., REPKO T., HOFFMAN E. J. THE EFFECT OF AMINO ACID DEPRIVATION ON SUBSEQUENT DEOXYRIBONUCLEIC ACID REPLICATION. Biochim Biophys Acta. 1963 Sep 17;76:9–24. [PubMed] [Google Scholar]
  30. 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]
  31. Light J., Molin S. Replication control functions of plasmid R1 act as inhibitors of expression of a gene required for replication. Mol Gen Genet. 1981;184(1):56–61. doi: 10.1007/BF00271195. [DOI] [PubMed] [Google Scholar]
  32. Løbner-Olesen A., Skarstad K., Hansen F. G., von Meyenburg K., Boye E. The DnaA protein determines the initiation mass of Escherichia coli K-12. Cell. 1989 Jun 2;57(5):881–889. doi: 10.1016/0092-8674(89)90802-7. [DOI] [PubMed] [Google Scholar]
  33. Monk M., Gross J. Induction of prophage lambda in a mutant of E. coli K12 defective in initiation of DNA replication at high temperature. Mol Gen Genet. 1971;110(4):299–306. doi: 10.1007/BF00438272. [DOI] [PubMed] [Google Scholar]
  34. Mulder E., Woldringh C. L. Actively replicating nucleoids influence positioning of division sites in Escherichia coli filaments forming cells lacking DNA. J Bacteriol. 1989 Aug;171(8):4303–4314. doi: 10.1128/jb.171.8.4303-4314.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Nordström K., Bernander R., Dasgupta S. Analysis of the bacterial cell cycle using strains in which chromosome replication is controlled by plasmid R1. Res Microbiol. 1991 Feb-Apr;142(2-3):181–188. doi: 10.1016/0923-2508(91)90028-9. [DOI] [PubMed] [Google Scholar]
  36. Nordström K., Bernander R., Dasgupta S. The Escherichia coli cell cycle: one cycle or multiple independent processes that are co-ordinated? Mol Microbiol. 1991 Apr;5(4):769–774. doi: 10.1111/j.1365-2958.1991.tb00747.x. [DOI] [PubMed] [Google Scholar]
  37. Nordström K., Wagner E. G. Kinetic aspects of control of plasmid replication by antisense RNA. Trends Biochem Sci. 1994 Jul;19(7):294–300. doi: 10.1016/0968-0004(94)90008-6. [DOI] [PubMed] [Google Scholar]
  38. Ogawa T., Okazaki T. Cell cycle-dependent transcription from the gid and mioC promoters of Escherichia coli. J Bacteriol. 1994 Mar;176(6):1609–1615. doi: 10.1128/jb.176.6.1609-1615.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Okada Y., Wachi M., Hirata A., Suzuki K., Nagai K., Matsuhashi M. Cytoplasmic axial filaments in Escherichia coli cells: possible function in the mechanism of chromosome segregation and cell division. J Bacteriol. 1994 Feb;176(3):917–922. doi: 10.1128/jb.176.3.917-922.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Pierucci O., Rickert M. Duplication of Escherichia coli during inhibition of net phospholipid synthesis. J Bacteriol. 1985 Apr;162(1):374–382. doi: 10.1128/jb.162.1.374-382.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. 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]
  42. Tang M. S., Helmstetter C. E. Coordination between chromosome replication and cell division in Escherichia coli. J Bacteriol. 1980 Mar;141(3):1148–1156. doi: 10.1128/jb.141.3.1148-1156.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Theisen P. W., Grimwade J. E., Leonard A. C., Bogan J. A., Helmstetter C. E. Correlation of gene transcription with the time of initiation of chromosome replication in Escherichia coli. Mol Microbiol. 1993 Nov;10(3):575–584. doi: 10.1111/j.1365-2958.1993.tb00929.x. [DOI] [PubMed] [Google Scholar]
  44. de Boer P. A., Cook W. R., Rothfield L. I. Bacterial cell division. Annu Rev Genet. 1990;24:249–274. doi: 10.1146/annurev.ge.24.120190.001341. [DOI] [PubMed] [Google Scholar]

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