Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1993 Jul;175(13):4016–4024. doi: 10.1128/jb.175.13.4016-4024.1993

Regulatory interactions between RepA, an essential replication protein, and the DNA repeats of RepFIB from plasmid P307.

A J Spiers 1, N Bhana 1, P L Bergquist 1
PMCID: PMC204830  PMID: 8320218

Abstract

The control of RepFIB replication appears to rely on the interaction between an initiator protein (RepA) and two sets of DNA repeat elements located on either side of the repA gene (BCDD'D" and EFGHIJ). In vivo genetic tests demonstrate that the BCDD'D" repeats form part of the origin of replication, while some of the downstream repeat elements (HIJ) are involved in the sensing and setting of plasmid copy number. RepA DNA binding to these groups of repeats has been investigated in vivo by utilizing the fact that the replicon contains three active promoters (orip, repAp, and EFp), one of which has previously been shown to control the expression of repA (repAp). All three promoters are closely associated with the repeat elements flanking repA, and an investigation using lacZ or cml gene fusions has demonstrated that RepA expressed in trans is able to repress each promoter. However, these assays suggest that the transcriptional responses of orip and repAp to RepA repression are significantly different, despite the fact that both promoters are embedded within the BCDD'D" repeat elements. Extra copies of the BCDD'D" or EFG repeats in trans have no effect on RepA repression of repAp embedded in a second copy of the BCDD'D" repeats, but copies of the HIJ or EFGHIJ repeats are able to derepress repAp, suggesting that there is a fundamental difference between RepA-BCDD'D" or -HIJ complexes and RepA-EFG or -EFGHIJ complexes.

Full text

PDF
4016

Images in this article

4021Image
on p.4021

Selected References

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

  1. ADELBERG E. A., BURNS S. N. Genetic variation in the sex factor of Escherichia coli. J Bacteriol. 1960 Mar;79:321–330. doi: 10.1128/jb.79.3.321-330.1960. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Abeles A. L., Austin S. J. Antiparallel plasmid-plasmid pairing may control P1 plasmid replication. Proc Natl Acad Sci U S A. 1991 Oct 15;88(20):9011–9015. doi: 10.1073/pnas.88.20.9011. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Abeles A. L., Austin S. J. P1 plasmid replication requires methylated DNA. EMBO J. 1987 Oct;6(10):3185–3189. doi: 10.1002/j.1460-2075.1987.tb02630.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Abeles A. L., Reaves L. D., Austin S. J. A single DnaA box is sufficient for initiation from the P1 plasmid origin. J Bacteriol. 1990 Aug;172(8):4386–4391. doi: 10.1128/jb.172.8.4386-4391.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bergquist P. L., Lane H. E., Malcolm L., Downard R. A. Molecular homology and incompatibility in the IncFI plasmid Group. J Gen Microbiol. 1982 Feb;128(2):223–238. doi: 10.1099/00221287-128-2-223. [DOI] [PubMed] [Google Scholar]
  6. Bramhill D., Kornberg A. Duplex opening by dnaA protein at novel sequences in initiation of replication at the origin of the E. coli chromosome. Cell. 1988 Mar 11;52(5):743–755. doi: 10.1016/0092-8674(88)90412-6. [DOI] [PubMed] [Google Scholar]
  7. Brendler T., Abeles A., Austin S. Critical sequences in the core of the P1 plasmid replication origin. J Bacteriol. 1991 Jul;173(13):3935–3942. doi: 10.1128/jb.173.13.3935-3942.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Brosius J. Plasmid vectors for the selection of promoters. Gene. 1984 Feb;27(2):151–160. doi: 10.1016/0378-1119(84)90136-7. [DOI] [PubMed] [Google Scholar]
  9. Chang A. C., Cohen S. N. Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. J Bacteriol. 1978 Jun;134(3):1141–1156. doi: 10.1128/jb.134.3.1141-1156.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Chattoraj D. K., Abeles A. L., Yarmolinsky M. B. P1 plasmid maintenance: a paradigm of precise control. Basic Life Sci. 1985;30:355–381. doi: 10.1007/978-1-4613-2447-8_27. [DOI] [PubMed] [Google Scholar]
  11. Chattoraj D., Cordes K., Abeles A. Plasmid P1 replication: negative control by repeated DNA sequences. Proc Natl Acad Sci U S A. 1984 Oct;81(20):6456–6460. doi: 10.1073/pnas.81.20.6456. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Fuller R. S., Kornberg A. Purified dnaA protein in initiation of replication at the Escherichia coli chromosomal origin of replication. Proc Natl Acad Sci U S A. 1983 Oct;80(19):5817–5821. doi: 10.1073/pnas.80.19.5817. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gammie A. E., Crosa J. H. Co-operative autoregulation of a replication protein gene. Mol Microbiol. 1991 Dec;5(12):3015–3023. doi: 10.1111/j.1365-2958.1991.tb01861.x. [DOI] [PubMed] [Google Scholar]
  14. Gammie A. E., Crosa J. H. Roles of DNA adenine methylation in controlling replication of the REPI replicon of plasmid pColV-K30. Mol Microbiol. 1991 Feb;5(2):495–503. doi: 10.1111/j.1365-2958.1991.tb02133.x. [DOI] [PubMed] [Google Scholar]
  15. Gardner R., McAnulty J., Feher E., Lane D. Location of rep and inc sequences in the F secondary replicon. Plasmid. 1985 Mar;13(2):145–148. doi: 10.1016/0147-619x(85)90067-8. [DOI] [PubMed] [Google Scholar]
  16. Hansen E. B., Yarmolinsky M. B. Host participation in plasmid maintenance: dependence upon dnaA of replicons derived from P1 and F. Proc Natl Acad Sci U S A. 1986 Jun;83(12):4423–4427. doi: 10.1073/pnas.83.12.4423. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Itoh Y., Terawaki Y. Replication properties of mini-Rts1 derivatives deleted for DnaA boxes in the replication origin. Plasmid. 1989 May;21(3):242–246. doi: 10.1016/0147-619x(89)90048-6. [DOI] [PubMed] [Google Scholar]
  18. Kogoma T., von Meyenburg K. The origin of replication, oriC, and the dnaA protein are dispensable in stable DNA replication (sdrA) mutants of Escherichia coli K-12. EMBO J. 1983;2(3):463–468. doi: 10.1002/j.1460-2075.1983.tb01445.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. LURIA S. E., BURROUS J. W. Hybridization between Escherichia coli and Shigella. J Bacteriol. 1957 Oct;74(4):461–476. doi: 10.1128/jb.74.4.461-476.1957. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Lederberg E. M., Cohen S. N. Transformation of Salmonella typhimurium by plasmid deoxyribonucleic acid. J Bacteriol. 1974 Sep;119(3):1072–1074. doi: 10.1128/jb.119.3.1072-1074.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Maas R., Saadi S., Maas W. K. Properties and incompatibility behavior of miniplasmids derived from the bireplicon plasmid pCG86. Mol Gen Genet. 1989 Aug;218(2):190–198. doi: 10.1007/BF00331268. [DOI] [PubMed] [Google Scholar]
  22. McEachern M. J., Bott M. A., Tooker P. A., Helinski D. R. Negative control of plasmid R6K replication: possible role of intermolecular coupling of replication origins. Proc Natl Acad Sci U S A. 1989 Oct;86(20):7942–7946. doi: 10.1073/pnas.86.20.7942. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Mukherjee S., Erickson H., Bastia D. Detection of DNA looping due to simultaneous interaction of a DNA-binding protein with two spatially separated binding sites on DNA. Proc Natl Acad Sci U S A. 1988 Sep;85(17):6287–6291. doi: 10.1073/pnas.85.17.6287. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Mukherjee S., Erickson H., Bastia D. Enhancer-origin interaction in plasmid R6K involves a DNA loop mediated by initiator protein. Cell. 1988 Feb 12;52(3):375–383. doi: 10.1016/s0092-8674(88)80030-8. [DOI] [PubMed] [Google Scholar]
  25. Mulligan M. E., Hawley D. K., Entriken R., McClure W. R. Escherichia coli promoter sequences predict in vitro RNA polymerase selectivity. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 2):789–800. doi: 10.1093/nar/12.1part2.789. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Nomura N., Masai H., Inuzuka M., Miyazaki C., Ohtsubo E., Itoh T., Sasamoto S., Matsui M., Ishizaki R., Arai K. Identification of eleven single-strand initiation sequences (ssi) for priming of DNA replication in the F, R6K, R100 and ColE2 plasmids. Gene. 1991 Dec 1;108(1):15–22. doi: 10.1016/0378-1119(91)90482-q. [DOI] [PubMed] [Google Scholar]
  27. Pal S. K., Chattoraj D. K. P1 plasmid replication: initiator sequestration is inadequate to explain control by initiator-binding sites. J Bacteriol. 1988 Aug;170(8):3554–3560. doi: 10.1128/jb.170.8.3554-3560.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Perez-Casal J. F., Crosa J. H. Novel incompatibility and partition loci for the REPI replication region of plasmid ColV-K30. J Bacteriol. 1987 Nov;169(11):5078–5086. doi: 10.1128/jb.169.11.5078-5086.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Perez-Casal J. F., Gammie A. E., Crosa J. H. Nucleotide sequence analysis and expression of the minimum REPI replication region and incompatibility determinants of pColV-K30. J Bacteriol. 1989 Apr;171(4):2195–2201. doi: 10.1128/jb.171.4.2195-2201.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Prentki P., Krisch H. M. In vitro insertional mutagenesis with a selectable DNA fragment. Gene. 1984 Sep;29(3):303–313. doi: 10.1016/0378-1119(84)90059-3. [DOI] [PubMed] [Google Scholar]
  31. Saul D., Spiers A. J., McAnulty J., Gibbs M. G., Bergquist P. L., Hill D. F. Nucleotide sequence and replication characteristics of RepFIB, a basic replicon of IncF plasmids. J Bacteriol. 1989 May;171(5):2697–2707. doi: 10.1128/jb.171.5.2697-2707.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Shultz J., Silhavy T. J., Berman M. L., Fiil N., Emr S. D. A previously unidentified gene in the spc operon of Escherichia coli K12 specifies a component of the protein export machinery. Cell. 1982 Nov;31(1):227–235. doi: 10.1016/0092-8674(82)90422-6. [DOI] [PubMed] [Google Scholar]
  33. Spiers A. J., Bergquist P. L. Expression and regulation of the RepA protein of the RepFIB replicon from plasmid P307. J Bacteriol. 1992 Dec;174(23):7533–7541. doi: 10.1128/jb.174.23.7533-7541.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Stalker D. M., McBride K. E., Malyj L. D. Herbicide resistance in transgenic plants expressing a bacterial detoxification gene. Science. 1988 Oct 21;242(4877):419–423. doi: 10.1126/science.242.4877.419. [DOI] [PubMed] [Google Scholar]
  35. Takeshita S., Sato M., Toba M., Masahashi W., Hashimoto-Gotoh T. High-copy-number and low-copy-number plasmid vectors for lacZ alpha-complementation and chloramphenicol- or kanamycin-resistance selection. Gene. 1987;61(1):63–74. doi: 10.1016/0378-1119(87)90365-9. [DOI] [PubMed] [Google Scholar]
  36. Trawick J. D., Kline B. C. A two-stage molecular model for control of mini-F replication. Plasmid. 1985 Jan;13(1):59–69. doi: 10.1016/0147-619x(85)90056-3. [DOI] [PubMed] [Google Scholar]
  37. Wickner S. H., Chattoraj D. K. Replication of mini-P1 plasmid DNA in vitro requires two initiation proteins, encoded by the repA gene of phage P1 and the dnaA gene of Escherichia coli. Proc Natl Acad Sci U S A. 1987 Jun;84(11):3668–3672. doi: 10.1073/pnas.84.11.3668. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Yang J., Pittard J. Molecular analysis of the regulatory region of the Escherichia coli K-12 tyrB gene. J Bacteriol. 1987 Oct;169(10):4710–4715. doi: 10.1128/jb.169.10.4710-4715.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. 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 Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES