Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1997 Apr;179(7):2163–2168. doi: 10.1128/jb.179.7.2163-2168.1997

Role of the RepA1 protein in RepFIC plasmid replication.

R Maas 1, C Wang 1
PMCID: PMC178951  PMID: 9079900

Abstract

Using a sensitive primer extension technique, we have carried out studies to localize the start site of replication of the replicon RepFIC. In the course of these studies, we have found evidence that supports the hypothesis that transcription is an integral component of the initiation of replication. On the basis of our findings, we suggest that the transcript is processed to act as a primer, and therefore we propose that the transcript has a dual role as primer of replication and mRNA for the RepA1 protein. We present a model, based on our evidence, for the initiation of replication of the replicon RepFIC. This model provides as well an alternative explanation for what has been called the cis action of RepA1, and we show that RepA1 may act in trans as well as in cis.

Full Text

The Full Text of this article is available as a PDF (733.8 KB).

Selected References

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

  1. Bernander R., Krabbe M., Nordström K. Mapping of the in vivo start site for leading strand DNA synthesis in plasmid R1. EMBO J. 1992 Dec;11(12):4481–4487. doi: 10.1002/j.1460-2075.1992.tb05549.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Blomberg P., Nordström K., Wagner E. G. Replication control of plasmid R1: RepA synthesis is regulated by CopA RNA through inhibition of leader peptide translation. EMBO J. 1992 Jul;11(7):2675–2683. doi: 10.1002/j.1460-2075.1992.tb05333.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Couturier M., Bex F., Bergquist P. L., Maas W. K. Identification and classification of bacterial plasmids. Microbiol Rev. 1988 Sep;52(3):375–395. doi: 10.1128/mr.52.3.375-395.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Dong X. N., Womble D. D., Rownd R. H. In-vivo studies on the cis-acting replication initiator protein of IncFII plasmid NR1. J Mol Biol. 1988 Aug 5;202(3):495–509. doi: 10.1016/0022-2836(88)90281-1. [DOI] [PubMed] [Google Scholar]
  5. Freitag M., Dighde N., Sachs M. S. A UV-induced mutation in neurospora that affects translational regulation in response to arginine. Genetics. 1996 Jan;142(1):117–127. doi: 10.1093/genetics/142.1.117. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hama C., Takizawa T., Moriwaki H., Urasaki Y., Mizobuchi K. Organization of the replication control region of plasmid ColIb-P9. J Bacteriol. 1990 Apr;172(4):1983–1991. doi: 10.1128/jb.172.4.1983-1991.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Itoh T., Tomizawa J. Formation of an RNA primer for initiation of replication of ColE1 DNA by ribonuclease H. Proc Natl Acad Sci U S A. 1980 May;77(5):2450–2454. doi: 10.1073/pnas.77.5.2450. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Lima T. M., Lim D. Isolation and characterization of host mutants defective in msDNA synthesis: role of ribonuclease H in msDNA synthesis. Plasmid. 1995 May;33(3):235–238. doi: 10.1006/plas.1995.1026. [DOI] [PubMed] [Google Scholar]
  9. Maas R., Oppenheim J., Saadi S., Fuchs T., Maas W. K. Isolation and properties of the RepA1 protein of the IncFII replicon, RepFIC. Mol Microbiol. 1991 Apr;5(4):927–932. doi: 10.1111/j.1365-2958.1991.tb00767.x. [DOI] [PubMed] [Google Scholar]
  10. Masai H., Arai K. Leading strand synthesis of R1 plasmid replication in vitro is primed by primase alone at a specific site downstream of oriR. J Biol Chem. 1989 May 15;264(14):8082–8090. [PubMed] [Google Scholar]
  11. Masai H., Arai K. RepA protein- and oriR-dependent initiation of R1 plasmid replication: identification of a rho-dependent transcription terminator required for cis-action of repA protein. Nucleic Acids Res. 1988 Jul 25;16(14A):6493–6514. doi: 10.1093/nar/16.14.6493. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Masai H., Kaziro Y., Arai K. Definition of oriR, the minimum DNA segment essential for initiation of R1 plasmid replication in vitro. Proc Natl Acad Sci U S A. 1983 Nov;80(22):6814–6818. doi: 10.1073/pnas.80.22.6814. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Miki T., Easton A. M., Rownd R. H. Cloning of replication, incompatibility, and stability functions of R plasmid NR1. J Bacteriol. 1980 Jan;141(1):87–99. doi: 10.1128/jb.141.1.87-99.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Miyazaki C., Kawai Y., Ohtsubo H., Ohtsubo E. Unidirectional replication of plasmid R100. J Mol Biol. 1988 Nov 20;204(2):331–343. doi: 10.1016/0022-2836(88)90580-3. [DOI] [PubMed] [Google Scholar]
  15. Nordström K., Molin S., Light J. Control of replication of bacterial plasmids: genetics, molecular biology, and physiology of the plasmid R1 system. Plasmid. 1984 Sep;12(2):71–90. doi: 10.1016/0147-619x(84)90054-4. [DOI] [PubMed] [Google Scholar]
  16. Picken R. N., Mazaitis A. J., Saadi S., Maas W. K. Characterization of the basic replicons of the chimeric R/Ent plasmid pCG86 and the related Ent plasmid P307. Plasmid. 1984 Jul;12(1):10–18. doi: 10.1016/0147-619x(84)90062-3. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. Saadi S., Maas W. K., Hill D. F., Bergquist P. L. Nucleotide sequence analysis of RepFIC, a basic replicon present in IncFI plasmids P307 and F, and its relation to the RepA replicon of IncFII plasmids. J Bacteriol. 1987 May;169(5):1836–1846. doi: 10.1128/jb.169.5.1836-1846.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. 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]
  20. Shiba K., Mizobuchi K. Posttranscriptional control of plasmid ColIb-P9 repZ gene expression by a small RNA. J Bacteriol. 1990 Apr;172(4):1992–1997. doi: 10.1128/jb.172.4.1992-1997.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Song H., Phillips S. E., Parsons M. R., Maas R. Crystallization and preliminary crystallographic analysis of RepA1, a replication control protein of the RepFIC replicon of enterotoxin plasmid EntP307. Proteins. 1996 May;25(1):137–138. doi: 10.1002/(SICI)1097-0134(199605)25:1<137::AID-PROT13>3.0.CO;2-L. [DOI] [PubMed] [Google Scholar]
  22. Tomizawa J., Itoh T. Plasmid ColE1 incompatibility determined by interaction of RNA I with primer transcript. Proc Natl Acad Sci U S A. 1981 Oct;78(10):6096–6100. doi: 10.1073/pnas.78.10.6096. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Wagner E. G., Simons R. W. Antisense RNA control in bacteria, phages, and plasmids. Annu Rev Microbiol. 1994;48:713–742. doi: 10.1146/annurev.mi.48.100194.003433. [DOI] [PubMed] [Google Scholar]
  24. Wilson I. W., Praszkier J., Pittard A. J. Molecular analysis of RNAI control of repB translation in IncB plasmids. J Bacteriol. 1994 Nov;176(21):6497–6508. doi: 10.1128/jb.176.21.6497-6508.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Womble D. D., Rownd R. H. Regulation of IncFII plasmid DNA replication. A quantitative model for control of plasmid NR1 replication in the bacterial cell division cycle. J Mol Biol. 1986 Dec 5;192(3):529–547. doi: 10.1016/0022-2836(86)90274-3. [DOI] [PubMed] [Google Scholar]
  26. Wu R., Wang X., Womble D. D., Rownd R. H. Expression of the repA1 gene of IncFII plasmid NR1 is translationally coupled to expression of an overlapping leader peptide. J Bacteriol. 1992 Dec;174(23):7620–7628. doi: 10.1128/jb.174.23.7620-7628.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES