Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1991 May 1;88(9):3867–3871. doi: 10.1073/pnas.88.9.3867

Replication terminator protein of Escherichia coli is a transcriptional repressor of its own synthesis.

S Natarajan 1, W L Kelley 1, D Bastia 1
PMCID: PMC51554  PMID: 2023933

Abstract

We have investigated the regulation of synthesis of the replication terminator protein (Ter) of Escherichia coli and have discovered that the protein is a repressor of its own synthesis at the transcriptional step. Since the synthesis of Ter protein was observed to be down-regulated in vivo, these results are consistent with autoregulation as one control mechanism of Ter protein within the cell. Analysis of the tus gene that encodes the Ter protein revealed that transcription was initiated from a single promoter located within the upstream nontranscribed sequence. In vitro footprinting experiments have revealed that Ter protein prevented binding of RNA polymerase to the promoter sequence when both proteins were incubated with promoter DNA. However, once bound to the promoter, RNA polymerase could not be displaced by Ter protein. Conversely, prebound Ter protein could not be dislodged from its binding site at the promoter when challenged with RNA polymerase. Therefore, Ter protein can serve as a transcriptional repressor of its own synthesis by preventing RNA polymerase from binding to the tus promoter when both proteins are present in the cell milieu.

Full text

PDF
3867

Images in this article

3868Image
on p.3868
3869Image
on p.3869
3869Image
on p.3869
3869Image
on p.3869
3870Image
on p.3870
3870Image
on p.3870
3870Image
on p.3870
3870Image
on p.3870

Selected References

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

  1. Aiba H., Adhya S., de Crombrugghe B. Evidence for two functional gal promoters in intact Escherichia coli cells. J Biol Chem. 1981 Nov 25;256(22):11905–11910. [PubMed] [Google Scholar]
  2. Atlung T., Clausen E. S., Hansen F. G. Autoregulation of the dnaA gene of Escherichia coli K12. Mol Gen Genet. 1985;200(3):442–450. doi: 10.1007/BF00425729. [DOI] [PubMed] [Google Scholar]
  3. Bastia D., Germino J., Crosa J. H., Ram J. The nucleotide sequence surrounding the replication terminus of R6K. Proc Natl Acad Sci U S A. 1981 Apr;78(4):2095–2099. doi: 10.1073/pnas.78.4.2095. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Braun R. E., O'Day K., Wright A. Autoregulation of the DNA replication gene dnaA in E. coli K-12. Cell. 1985 Jan;40(1):159–169. doi: 10.1016/0092-8674(85)90319-8. [DOI] [PubMed] [Google Scholar]
  5. Brewer B. J., Fangman W. L. A replication fork barrier at the 3' end of yeast ribosomal RNA genes. Cell. 1988 Nov 18;55(4):637–643. doi: 10.1016/0092-8674(88)90222-x. [DOI] [PubMed] [Google Scholar]
  6. Burgess R. R., Jendrisak J. J. A procedure for the rapid, large-scall purification of Escherichia coli DNA-dependent RNA polymerase involving Polymin P precipitation and DNA-cellulose chromatography. Biochemistry. 1975 Oct 21;14(21):4634–4638. doi: 10.1021/bi00692a011. [DOI] [PubMed] [Google Scholar]
  7. Cammack K. A., Wade H. E. The sedimentation behaviour of ribonuclease-active and -inactive ribosomes from bacteria. Biochem J. 1965 Sep;96(3):671–680. doi: 10.1042/bj0960671. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Crosa J. H., Luttropp L. K., Falkow S. Mode of replication of the conjugative R-plasmid RSF1040 in Escherichia coli. J Bacteriol. 1976 Apr;126(1):454–466. doi: 10.1128/jb.126.1.454-466.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Davison B. L., Leighton T., Rabinowitz J. C. Purification of Bacillus subtilis RNA polymerase with heparin-agarose. In vitro transcription of phi 29 DNA. J Biol Chem. 1979 Sep 25;254(18):9220–9226. [PubMed] [Google Scholar]
  10. Donachie W. D., Begg K. J. Chromosome partition in Escherichia coli requires postreplication protein synthesis. J Bacteriol. 1989 Oct;171(10):5405–5409. doi: 10.1128/jb.171.10.5405-5409.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Filutowicz M., Davis G., Greener A., Helinski D. R. Autorepressor properties of the pi-initiation protein encoded by plasmid R6K. Nucleic Acids Res. 1985 Jan 11;13(1):103–114. doi: 10.1093/nar/13.1.103. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Germino J., Bastia D. Termination of DNA replication in vitro at a sequence-specific replication terminus. Cell. 1981 Mar;23(3):681–687. doi: 10.1016/0092-8674(81)90431-1. [DOI] [PubMed] [Google Scholar]
  13. Grossman N., Rosner E., Ron E. Z. Termination of DNA replication is required for cell division in Escherichia coli. J Bacteriol. 1989 Jan;171(1):74–79. doi: 10.1128/jb.171.1.74-79.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Henson J. M., Kuempel P. L. Deletion of the terminus region (340 kilobase pairs of DNA) from the chromosome of Escherichia coli. Proc Natl Acad Sci U S A. 1985 Jun;82(11):3766–3770. doi: 10.1073/pnas.82.11.3766. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hill T. M., Marians K. J. Escherichia coli Tus protein acts to arrest the progression of DNA replication forks in vitro. Proc Natl Acad Sci U S A. 1990 Apr;87(7):2481–2485. doi: 10.1073/pnas.87.7.2481. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hill T. M., Tecklenburg M. L., Pelletier A. J., Kuempel P. L. tus, the trans-acting gene required for termination of DNA replication in Escherichia coli, encodes a DNA-binding protein. Proc Natl Acad Sci U S A. 1989 Mar;86(5):1593–1597. doi: 10.1073/pnas.86.5.1593. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Horiuchi T., Hidaka M. Core sequence of two separable terminus sites of the R6K plasmid that exhibit polar inhibition of replication is a 20 bp inverted repeat. Cell. 1988 Aug 12;54(4):515–523. doi: 10.1016/0092-8674(88)90073-6. [DOI] [PubMed] [Google Scholar]
  18. Kelley W., Bastia D. Replication initiator protein of plasmid R6K autoregulates its own synthesis at the transcriptional step. Proc Natl Acad Sci U S A. 1985 May;82(9):2574–2578. doi: 10.1073/pnas.82.9.2574. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Khatri G. S., MacAllister T., Sista P. R., Bastia D. The replication terminator protein of E. coli is a DNA sequence-specific contra-helicase. Cell. 1989 Nov 17;59(4):667–674. doi: 10.1016/0092-8674(89)90012-3. [DOI] [PubMed] [Google Scholar]
  20. Kolter R., Helinski D. R. Activity of the replication terminus of plasmid R6K in hybrid replicons in Escherichia coli. J Mol Biol. 1978 Sep 25;124(3):425–441. doi: 10.1016/0022-2836(78)90180-8. [DOI] [PubMed] [Google Scholar]
  21. 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]
  22. Lee E. H., Kornberg A., Hidaka M., Kobayashi T., Horiuchi T. Escherichia coli replication termination protein impedes the action of helicases. Proc Natl Acad Sci U S A. 1989 Dec;86(23):9104–9108. doi: 10.1073/pnas.86.23.9104. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. 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]
  24. MacAllister T., Khatri G. S., Bastia D. Sequence-specific and polarized replication termination in vitro: complementation of extracts of tus- Escherichia coli by purified Ter protein and analysis of termination intermediates. Proc Natl Acad Sci U S A. 1990 Apr;87(7):2828–2832. doi: 10.1073/pnas.87.7.2828. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Maxam A. M., Gilbert W. A new method for sequencing DNA. Proc Natl Acad Sci U S A. 1977 Feb;74(2):560–564. doi: 10.1073/pnas.74.2.560. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Simons R. W., Houman F., Kleckner N. Improved single and multicopy lac-based cloning vectors for protein and operon fusions. Gene. 1987;53(1):85–96. doi: 10.1016/0378-1119(87)90095-3. [DOI] [PubMed] [Google Scholar]
  27. Sista P. R., Hutchinson C. A., 3rd, Bastia D. DNA-protein interaction at the replication termini of plasmid R6K. Genes Dev. 1991 Jan;5(1):74–82. doi: 10.1101/gad.5.1.74. [DOI] [PubMed] [Google Scholar]
  28. Sista P. R., Mukherjee S., Patel P., Khatri G. S., Bastia D. A host-encoded DNA-binding protein promotes termination of plasmid replication at a sequence-specific replication terminus. Proc Natl Acad Sci U S A. 1989 May;86(9):3026–3030. doi: 10.1073/pnas.86.9.3026. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Vocke C., Bastia D. The replication initiator protein of plasmid pSC101 is a transcriptional repressor of its own cistron. Proc Natl Acad Sci U S A. 1985 Apr;82(8):2252–2256. doi: 10.1073/pnas.82.8.2252. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Yamaguchi K., Masamune Y. Autogenous regulation of synthesis of the replication protein in plasmid pSC101. Mol Gen Genet. 1985;200(3):362–367. doi: 10.1007/BF00425718. [DOI] [PubMed] [Google Scholar]
  31. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES