Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1997 May;179(10):3358–3361. doi: 10.1128/jb.179.10.3358-3361.1997

Search for additional replication terminators in the Bacillus subtilis 168 chromosome.

A A Griffiths 1, R G Wake 1
PMCID: PMC179119  PMID: 9150236

Abstract

The Bacillus subtilis 168 chromosome is known to contain at least six DNA replication terminators in the terminus region of the chromosome. By using a degenerate DNA probe for the consensus terminator sequence and low-stringency hybridization conditions, several additional minor hybridizing bands were identified. DNA corresponding to the most intense of these bands was cloned and characterized. Although localized in the terminus region, it could not bind RTP and possibly represents a degenerate terminator. A search of the SubtiList database identified an additional terminator sequence in the terminus region, near glnA. It was shown to bind RTP and to function in blocking replication fork movement in a polar manner. Its orientation conformed to the replication fork trap arrangement of the other terminators. The low-stringency hybridization experiments failed to identify any terminus region-type terminators in the region of the chromosome where postinitiation control sequences (STer sites) are known to reside. The two most likely terminators in STer site regions, in terms of sequence similarity to terminus region terminators, were identified through sequence searching. They were synthesized and were found not to bind RTP under conditions that allowed binding to terminus region terminators. Neither did they elicit fork arrest, when present in a plasmid, under stringent conditions. It is concluded that the STer site terminators, at least the first two to the left of oriC, do not have the typical consensus A+B site makeup of terminus region terminators.

Full Text

The Full Text of this article is available as a PDF (272.2 KB).

Selected References

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

  1. Azevedo V., Alvarez E., Zumstein E., Damiani G., Sgaramella V., Ehrlich S. D., Serror P. An ordered collection of Bacillus subtilis DNA segments cloned in yeast artificial chromosomes. Proc Natl Acad Sci U S A. 1993 Jul 1;90(13):6047–6051. doi: 10.1073/pnas.90.13.6047. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. 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]
  3. 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]
  4. François V., Louarn J., Louarn J. M. The terminus of the Escherichia coli chromosome is flanked by several polar replication pause sites. Mol Microbiol. 1989 Aug;3(8):995–1002. doi: 10.1111/j.1365-2958.1989.tb00250.x. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. Kuempel P. L., Pelletier A. J., Hill T. M. Tus and the terminators: the arrest of replication in prokaryotes. Cell. 1989 Nov 17;59(4):581–583. doi: 10.1016/0092-8674(89)90001-9. [DOI] [PubMed] [Google Scholar]
  7. Langley D. B., Smith M. T., Lewis P. J., Wake R. G. Protein-nucleoside contacts in the interaction between the replication terminator protein of Bacillus subtilis and the DNA terminator. Mol Microbiol. 1993 Nov;10(4):771–779. doi: 10.1111/j.1365-2958.1993.tb00947.x. [DOI] [PubMed] [Google Scholar]
  8. Levine A., Autret S., Séror S. J. A checkpoint involving RTP, the replication terminator protein, arrests replication downstream of the origin during the Stringent Response in Bacillus subtilis. Mol Microbiol. 1995 Jan;15(2):287–295. doi: 10.1111/j.1365-2958.1995.tb02243.x. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. Lewis P. J., Wake R. G. DNA and protein sequence conservation at the replication terminus in Bacillus subtilis 168 and W23. J Bacteriol. 1989 Mar;171(3):1402–1408. doi: 10.1128/jb.171.3.1402-1408.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Meijer W. J., Smith M., Wake R. G., de Boer A. L., Venema G., Bron S. Identification and characterization of a novel type of replication terminator with bidirectional activity on the Bacillus subtilis theta plasmid pLS20. Mol Microbiol. 1996 Mar;19(6):1295–1306. doi: 10.1111/j.1365-2958.1996.tb02474.x. [DOI] [PubMed] [Google Scholar]
  12. Mohanty B. K., Sahoo T., Bastia D. The relationship between sequence-specific termination of DNA replication and transcription. EMBO J. 1996 May 15;15(10):2530–2539. [PMC free article] [PubMed] [Google Scholar]
  13. Moszer I., Glaser P., Danchin A. SubtiList: a relational database for the Bacillus subtilis genome. Microbiology. 1995 Feb;141(Pt 2):261–268. doi: 10.1099/13500872-141-2-261. [DOI] [PubMed] [Google Scholar]
  14. Smith M. T., Langley D. B., Young P. A., Wake R. G. The minimal sequence needed to define a functional DNA terminator in Bacillus subtilis. J Mol Biol. 1994 Aug 19;241(3):335–340. doi: 10.1006/jmbi.1994.1510. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. Smith M. T., de Vries C. J., Langley D. B., King G. F., Wake R. G. The Bacillus subtilis DNA replication terminator. J Mol Biol. 1996 Jul 5;260(1):54–69. doi: 10.1006/jmbi.1996.0381. [DOI] [PubMed] [Google Scholar]
  17. Strauch M. A., Aronson A. I., Brown S. W., Schreier H. J., Sonenhein A. L. Sequence of the Bacillus subtilis glutamine synthetase gene region. Gene. 1988 Nov 30;71(2):257–265. doi: 10.1016/0378-1119(88)90042-x. [DOI] [PubMed] [Google Scholar]
  18. Wake R., King G. A tale of two terminators: crystal structures sharpen the debate on DNA replication fork arrest mechanisms. Structure. 1997 Jan 15;5(1):1–5. doi: 10.1016/s0969-2126(97)00160-3. [DOI] [PubMed] [Google Scholar]

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

RESOURCES