Skip to main content
NCBI home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1992 Jan 11;20(1):105–109. doi: 10.1093/nar/20.1.105

Inhibition of DNA synthesis at the hemimethylated pBR322 origin of replication by a cell membrane fraction.

A Malki 1, R Kern 1, M Kohiyama 1, P Hughes 1
PMCID: PMC310332  PMID: 1738587

Abstract

The replication of both ColE1-type plasmids and plasmids bearing the origin of replication of the Escherichia coli chromosome (oriC) has been shown to be inhibited by hemimethylation of adenine residues within GATC sequences. In the case of oriC plasmids, this inhibition was previously shown to be mediated by the specific affinity of the hemimethylated origin DNA for an outer cell membrane fraction. Here, we suggest that a similar mechanism is operating in the case of the ColE1-like plasmid pBR322 as (i) a hemimethylated DNA fragment carrying the promoter for the RNA which primes DNA synthesis (RNAII) is specifically bound by the same membrane fraction and, (ii) the addition of the membrane fraction to a soluble assay of pBR322 replication results in preferential inhibition of initiation on the hemimethylated template. We suggest that membrane sequestration of hemimethylated origin DNA and/or associated replication genes following replication may be a common element restricting DNA replication to precise moments in the cell cycle.

Full text

PDF
105

Images in this article

106Image
on p.106
107Image
on p.107

Selected References

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

  1. Bakker A., Smith D. W. Methylation of GATC sites is required for precise timing between rounds of DNA replication in Escherichia coli. J Bacteriol. 1989 Oct;171(10):5738–5742. doi: 10.1128/jb.171.10.5738-5742.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bazaral M., Helinski D. R. Replication of a bacterial plasmid and an episome in Escherichia coli. Biochemistry. 1970 Jan 20;9(2):399–406. doi: 10.1021/bi00804a029. [DOI] [PubMed] [Google Scholar]
  3. Boye E., Løbner-Olesen A. The role of dam methyltransferase in the control of DNA replication in E. coli. Cell. 1990 Sep 7;62(5):981–989. doi: 10.1016/0092-8674(90)90272-g. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. Chan P. T., Ohmori H., Tomizawa J., Lebowitz J. Nucleotide sequence and gene organization of ColE1 DNA. J Biol Chem. 1985 Jul 25;260(15):8925–8935. [PubMed] [Google Scholar]
  6. Fuller R. S., Kaguni J. M., Kornberg A. Enzymatic replication of the origin of the Escherichia coli chromosome. Proc Natl Acad Sci U S A. 1981 Dec;78(12):7370–7374. doi: 10.1073/pnas.78.12.7370. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hughes P., Landoulsi A., Kohiyama M. A novel role for cAMP in the control of the activity of the E. coli chromosome replication initiator protein, DnaA. Cell. 1988 Oct 21;55(2):343–350. doi: 10.1016/0092-8674(88)90057-8. [DOI] [PubMed] [Google Scholar]
  8. Hughes P., Squali-Houssaini F., Forterre P., Kohiyama M. In vitro replication of a dam methylated and non-methylated ori-C plasmid. J Mol Biol. 1984 Jun 15;176(1):155–159. doi: 10.1016/0022-2836(84)90386-3. [DOI] [PubMed] [Google Scholar]
  9. Landoulsi A., Hughes P., Kern R., Kohiyama M. dam methylation and the initiation of DNA replication on oriC plasmids. Mol Gen Genet. 1989 Apr;216(2-3):217–223. doi: 10.1007/BF00334359. [DOI] [PubMed] [Google Scholar]
  10. Landoulsi A., Malki A., Kern R., Kohiyama M., Hughes P. The E. coli cell surface specifically prevents the initiation of DNA replication at oriC on hemimethylated DNA templates. Cell. 1990 Nov 30;63(5):1053–1060. doi: 10.1016/0092-8674(90)90508-c. [DOI] [PubMed] [Google Scholar]
  11. Messer W., Bellekes U., Lother H. Effect of dam methylation on the activity of the E. coli replication origin, oriC. EMBO J. 1985 May;4(5):1327–1332. doi: 10.1002/j.1460-2075.1985.tb03780.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Messer W., Noyer-Weidner M. Timing and targeting: the biological functions of Dam methylation in E. coli. Cell. 1988 Sep 9;54(6):735–737. doi: 10.1016/s0092-8674(88)90911-7. [DOI] [PubMed] [Google Scholar]
  13. Nordström K. Replication of plasmid R1: Meselson-Stahl density shift experiments revisited. Plasmid. 1983 Mar;9(2):218–221. doi: 10.1016/0147-619x(83)90023-9. [DOI] [PubMed] [Google Scholar]
  14. Ogden G. B., Pratt M. J., Schaechter M. The replicative origin of the E. coli chromosome binds to cell membranes only when hemimethylated. Cell. 1988 Jul 1;54(1):127–135. doi: 10.1016/0092-8674(88)90186-9. [DOI] [PubMed] [Google Scholar]
  15. Patnaik P. K., Merlin S., Polisky B. Effect of altering GATC sequences in the plasmid ColE1 primer promoter. J Bacteriol. 1990 Apr;172(4):1762–1768. doi: 10.1128/jb.172.4.1762-1768.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Russell D. W., Zinder N. D. Hemimethylation prevents DNA replication in E. coli. Cell. 1987 Sep 25;50(7):1071–1079. doi: 10.1016/0092-8674(87)90173-5. [DOI] [PubMed] [Google Scholar]
  17. Smith D. W., Garland A. M., Herman G., Enns R. E., Baker T. A., Zyskind J. W. Importance of state of methylation of oriC GATC sites in initiation of DNA replication in Escherichia coli. EMBO J. 1985 May;4(5):1319–1326. doi: 10.1002/j.1460-2075.1985.tb03779.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Sparks R. B., Jr, Helinski D. R. Association of cellular membrane of E. coli minicells with the origins/terminus region of replication of plasmid ColEl DNA. Nature. 1979 Feb 15;277(5697):572–574. doi: 10.1038/277572a0. [DOI] [PubMed] [Google Scholar]
  19. Sutcliffe J. G. Complete nucleotide sequence of the Escherichia coli plasmid pBR322. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 1):77–90. doi: 10.1101/sqb.1979.043.01.013. [DOI] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES