Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1996 Jul;178(14):4258–4265. doi: 10.1128/jb.178.14.4258-4265.1996

Replication fork arrest at relocated replication terminators on the Bacillus subtilis chromosome.

A H Franks 1, R G Wake 1
PMCID: PMC178184  PMID: 8763955

Abstract

The replication terminus region of the Bacillus subtilis chromosome, comprising TerI and TerII plus the rtp gene (referred to as the terC region) was relocated to serC (257 degrees) and cym (10 degrees) on the anticlockwise- and clockwise-replicating segments of the chromosome, respectively. In both cases, it was found that only the orientation of the terC region that placed TerI in opposition to the approaching replication fork was functional in fork arrest. When TerII was opposed to the approaching fork, it was nonfunctional. These findings confirm and extend earlier work which involved relocations to only the clockwise-replicating segment, at metD (100 degrees) and pyr (139 degrees). In the present work, it was further shown that in the strain in which TerII was opposed to an approaching fork at metD, overproduction of the replication terminator protein (RTP) enabled TerII to function as an arrest site. Thus, chromosomal TerII is nonfunctional in arrest in vivo because of a limiting level of RTP. Marker frequency analysis showed that TerI at both cym and metD caused only transient arrest of a replication fork. Arrest appeared to be more severe in the latter situation and caused the two forks to meet at approximately 145 degrees (just outside or on the edge of the replication fork trap). The minimum pause time erected by TerI at metD was calculated to be approximately 40% of the time taken to complete a round of replication. This significant pause at metD caused the cells to become elongated, indicating that cell division was delayed. Further work is needed to establish the immediate cause of the delay in division.

Full Text

The Full Text of this article is available as a PDF (559.2 KB).

Selected References

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

  1. Ahn K. S., Malo M. S., Smith M. T., Wake R. G. Autoregulation of the gene encoding the replication terminator protein of Bacillus subtilis. Gene. 1993 Sep 30;132(1):7–13. doi: 10.1016/0378-1119(93)90508-z. [DOI] [PubMed] [Google Scholar]
  2. Baker T. A. Replication arrest. Cell. 1995 Feb 24;80(4):521–524. doi: 10.1016/0092-8674(95)90504-9. [DOI] [PubMed] [Google Scholar]
  3. Carrigan C. M., Haarsma J. A., Smith M. T., Wake R. G. Sequence features of the replication terminus of the Bacillus subtilis chromosome. Nucleic Acids Res. 1987 Oct 26;15(20):8501–8509. doi: 10.1093/nar/15.20.8501. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Carrigan C. M., Pack R. A., Smith M. T., Wake R. G. Normal terC-region of the Bacillus subtilis chromosome acts in a polar manner to arrest the clockwise replication fork. J Mol Biol. 1991 Nov 20;222(2):197–207. doi: 10.1016/0022-2836(91)90206-l. [DOI] [PubMed] [Google Scholar]
  5. Franks A. H., Griffiths A. A., Wake R. G. Identification and characterization of new DNA replication terminators in Bacillus subtilis. Mol Microbiol. 1995 Jul;17(1):13–23. doi: 10.1111/j.1365-2958.1995.mmi_17010013.x. [DOI] [PubMed] [Google Scholar]
  6. Hauser P. M., Errington J. Characterization of cell cycle events during the onset of sporulation in Bacillus subtilis. J Bacteriol. 1995 Jul;177(14):3923–3931. doi: 10.1128/jb.177.14.3923-3931.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hill T. M. Arrest of bacterial DNA replication. Annu Rev Microbiol. 1992;46:603–633. doi: 10.1146/annurev.mi.46.100192.003131. [DOI] [PubMed] [Google Scholar]
  8. Horiuchi T., Fujimura Y. Recombinational rescue of the stalled DNA replication fork: a model based on analysis of an Escherichia coli strain with a chromosome region difficult to replicate. J Bacteriol. 1995 Feb;177(3):783–791. doi: 10.1128/jb.177.3.783-791.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Iismaa T. P., Smith M. T., Wake R. G. Physical map of the Bacillus subtilis replication terminus region: its confirmation, extension and genetic orientation. Gene. 1984 Dec;32(1-2):171–180. doi: 10.1016/0378-1119(84)90045-3. [DOI] [PubMed] [Google Scholar]
  10. Lewis P. J., Ralston G. B., Christopherson R. I., Wake R. G. Identification of the replication terminator protein binding sites in the terminus region of the Bacillus subtilis chromosome and stoichiometry of the binding. J Mol Biol. 1990 Jul 5;214(1):73–84. doi: 10.1016/0022-2836(90)90147-E. [DOI] [PubMed] [Google Scholar]
  11. Lewis P. J., Smith M. T., Wake R. G. A protein involved in termination of chromosome replication in Bacillus subtilis binds specifically to the terC site. J Bacteriol. 1989 Jun;171(6):3564–3567. doi: 10.1128/jb.171.6.3564-3567.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. McGinness T., Wake R. G. Division septation in the absence of chromosome termination in Bacillus subtilis. J Mol Biol. 1979 Oct 25;134(2):251–264. doi: 10.1016/0022-2836(79)90035-4. [DOI] [PubMed] [Google Scholar]
  13. Schneider A. M., Gaisne M., Anagnostopoulos C. Genetic structure and internal rearrangements of stable merodiploids from Bacillus subtilis strains carrying the trpE26 mutation. Genetics. 1982 Jun;101(2):189–210. doi: 10.1093/genetics/101.2.189. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Setlow B., Magill N., Febbroriello P., Nakhimovsky L., Koppel D. E., Setlow P. Condensation of the forespore nucleoid early in sporulation of Bacillus species. J Bacteriol. 1991 Oct;173(19):6270–6278. doi: 10.1128/jb.173.19.6270-6278.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Sharma B., Hill T. M. Insertion of inverted Ter sites into the terminus region of the Escherichia coli chromosome delays completion of DNA replication and disrupts the cell cycle. Mol Microbiol. 1995 Oct;18(1):45–61. doi: 10.1111/j.1365-2958.1995.mmi_18010045.x. [DOI] [PubMed] [Google Scholar]
  16. Smith M. T., Wake R. G. Definition and polarity of action of DNA replication terminators in Bacillus subtilis. J Mol Biol. 1992 Oct 5;227(3):648–657. doi: 10.1016/0022-2836(92)90214-5. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. Weiss A. S., Wake R. G. Restriction map of DNA spanning the replication terminus of the Bacillus subtilis chromosome. J Mol Biol. 1983 Dec 5;171(2):119–137. doi: 10.1016/s0022-2836(83)80349-0. [DOI] [PubMed] [Google Scholar]
  19. Wu L. J., Franks A. H., Wake R. G. Replication through the terminus region of the Bacillus subtilis chromosome is not essential for the formation of a division septum that partitions the DNA. J Bacteriol. 1995 Oct;177(19):5711–5715. doi: 10.1128/jb.177.19.5711-5715.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES