Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1988 Sep;170(9):4293–4298. doi: 10.1128/jb.170.9.4293-4298.1988

Location of sites that inhibit progression of replication forks in the terminus region of Escherichia coli.

A J Pelletier 1, T M Hill 1, P L Kuempel 1
PMCID: PMC211440  PMID: 2842310

Abstract

We used a Southern hybridization assay to locate precisely the sites at which DNA replication is arrested in the terminus region of the Escherichia coli chromosome. The assay was based on the properties of restriction fragments that contain stalled replication forks. Replication forks that entered the terminus from the clockwise direction with respect to the genetic map were inhibited near manA at a site called T2, which we located at kilobase 442 on the physical map of Bouché (J. P. Bouché, J. Mol. Biol. 154:1-20, 1982). Those that entered the terminus region traveling in the counterclockwise direction were inhibited near pyrF at a site called T1, which we located at kilobase 90. In each case we found only a single, precise site of arrest. Inhibition at T1 was not detectable in our assay in strains lacking the trans-acting locus tus, which is located near T2 and is required for T1 to function. We demonstrated that the sites of inhibition are also used during termination of replication in exponentially growing, wild-type cells. In all previous studies on the terminus of E. coli, inhibition has only been detected in strains that were modified so that the origin used was placed near the terminus to force the use of the sites of inhibition.

Full text

PDF
4297

Images in this article

4295Image
on p.4295
4295Image
on p.4295
4296Image
on p.4296
4296Image
on p.4296

Selected References

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

  1. Bachmann B. J. Linkage map of Escherichia coli K-12, edition 7. Microbiol Rev. 1983 Jun;47(2):180–230. doi: 10.1128/mr.47.2.180-230.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Baran N., Lapidot A., Manor H. Unusual sequence element found at the end of an amplicon. Mol Cell Biol. 1987 Jul;7(7):2636–2640. doi: 10.1128/mcb.7.7.2636. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bastia D., Germino J., Crosa J. H., Hale P. Molecular cloning of the replication terminus of the plasmid R6K. Gene. 1981 Jun-Jul;14(1-2):81–89. doi: 10.1016/0378-1119(81)90150-5. [DOI] [PubMed] [Google Scholar]
  4. Bouché J. P. Physical map of a 470 x 10(3) base-pair region flanking the terminus of DNA replication in the Escherichia coli K12 genome. J Mol Biol. 1982 Jan 5;154(1):1–20. doi: 10.1016/0022-2836(82)90413-2. [DOI] [PubMed] [Google Scholar]
  5. Gourse R. L., Takebe Y., Sharrock R. A., Nomura M. Feedback regulation of rRNA and tRNA synthesis and accumulation of free ribosomes after conditional expression of rRNA genes. Proc Natl Acad Sci U S A. 1985 Feb;82(4):1069–1073. doi: 10.1073/pnas.82.4.1069. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hanley P. J., Carrigan C. M., Rowe D. B., Wake R. G. Breakdown and quantitation of the forked termination of replication intermediate of Bacillus subtilis. J Mol Biol. 1987 Aug 5;196(3):721–727. doi: 10.1016/0022-2836(87)90043-x. [DOI] [PubMed] [Google Scholar]
  7. Hernández P., Lamm S. S., Bjerknes C. A., Hof J. V. Replication termini in the rDNA of synchronized pea root cells (Pisum sativum). EMBO J. 1988 Feb;7(2):303–308. doi: 10.1002/j.1460-2075.1988.tb02813.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hill C. W., Harnish B. W. Inversions between ribosomal RNA genes of Escherichia coli. Proc Natl Acad Sci U S A. 1981 Nov;78(11):7069–7072. doi: 10.1073/pnas.78.11.7069. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hill T. M., Henson J. M., Kuempel P. L. The terminus region of the Escherichia coli chromosome contains two separate loci that exhibit polar inhibition of replication. Proc Natl Acad Sci U S A. 1987 Apr;84(7):1754–1758. doi: 10.1073/pnas.84.7.1754. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hill T. M., Kopp B. J., Kuempel P. L. Termination of DNA replication in Escherichia coli requires a trans-acting factor. J Bacteriol. 1988 Feb;170(2):662–668. doi: 10.1128/jb.170.2.662-668.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kohara Y., Akiyama K., Isono K. The physical map of the whole E. coli chromosome: application of a new strategy for rapid analysis and sorting of a large genomic library. Cell. 1987 Jul 31;50(3):495–508. doi: 10.1016/0092-8674(87)90503-4. [DOI] [PubMed] [Google Scholar]
  12. Kuempel P. L., Duerr S. A., Seeley N. R. Terminus region of the chromosome in Escherichia coli inhibits replication forks. Proc Natl Acad Sci U S A. 1977 Sep;74(9):3927–3931. doi: 10.1073/pnas.74.9.3927. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Louarn J., Patte J., Louarn J. M. Evidence for a fixed termination site of chromosome replication in Escherichia coli K12. J Mol Biol. 1977 Sep 25;115(3):295–314. doi: 10.1016/0022-2836(77)90156-5. [DOI] [PubMed] [Google Scholar]
  14. Weiss A. S., Wake R. G. A unique DNA intermediate associated with termination of chromosome replication in Bacillus subtilis. Cell. 1984 Dec;39(3 Pt 2):683–689. doi: 10.1016/0092-8674(84)90475-6. [DOI] [PubMed] [Google Scholar]
  15. Weiss A. S., Wake R. G. Impediment to replication fork movement in the terminus region of the Bacillus subtilis chromosome. J Mol Biol. 1984 Nov 15;179(4):745–750. doi: 10.1016/0022-2836(84)90165-7. [DOI] [PubMed] [Google Scholar]
  16. de Massy B., Béjar S., Louarn J., Louarn J. M., Bouché J. P. Inhibition of replication forks exiting the terminus region of the Escherichia coli chromosome occurs at two loci separated by 5 min. Proc Natl Acad Sci U S A. 1987 Apr;84(7):1759–1763. doi: 10.1073/pnas.84.7.1759. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES