Skip to main content
PMC 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
. 1985 May;82(9):2588–2592. doi: 10.1073/pnas.82.9.2588

P1 plasmid replication: multiple functions of RepA protein at the origin.

D K Chattoraj, K M Snyder, A L Abeles
PMCID: PMC397609  PMID: 3857601

Abstract

Replication functions of a bacteriophage P1 miniplasmid are carried on a 1.2-kilobase pair (kb) segment that can be subdivided into a 245-base pair (bp) replication origin and a 959-bp region that encodes a protein required for replication (RepA). The origin region contains five 19-bp direct repeats. By using primer extension and gene-fusion assays, we mapped the promoter of the repA gene within the repeated sequences and showed that the promoter is repressed by RepA. Regulation of RepA synthesis is apparently achieved by the binding of RepA to the repeat sequences. This regulation might be a key step in the replication-control circuit, as we found that overproduction of RepA (from a foreign promoter) inhibits replication. Thus, in addition to being an autoregulated activator of replication, the protein also can have a negative regulatory role.

Full text

PDF

Images in this article

2589Image
on p.2589
2591Image
on p.2591

Selected References

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

  1. Abeles A. L., Snyder K. M., Chattoraj D. K. P1 plasmid replication: replicon structure. J Mol Biol. 1984 Mar 5;173(3):307–324. doi: 10.1016/0022-2836(84)90123-2. [DOI] [PubMed] [Google Scholar]
  2. 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]
  3. Baumstark B. R., Lowery K., Scott J. R. Location by DNA sequence analysis of cop mutations affecting the number of plasmid copies of prophage P1. Mol Gen Genet. 1984;194(3):513–516. doi: 10.1007/BF00425567. [DOI] [PubMed] [Google Scholar]
  4. Benyajati C., Dray J. F. Cloned Drosophila alcohol dehydrogenase genes are correctly expressed after transfection into Drosophila cells in culture. Proc Natl Acad Sci U S A. 1984 Mar;81(6):1701–1705. doi: 10.1073/pnas.81.6.1701. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bremer E., Silhavy T. J., Weisemann J. M., Weinstock G. M. Lambda placMu: a transposable derivative of bacteriophage lambda for creating lacZ protein fusions in a single step. J Bacteriol. 1984 Jun;158(3):1084–1093. doi: 10.1128/jb.158.3.1084-1093.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Brosius J., Dull T. J., Sleeter D. D., Noller H. F. Gene organization and primary structure of a ribosomal RNA operon from Escherichia coli. J Mol Biol. 1981 May 15;148(2):107–127. doi: 10.1016/0022-2836(81)90508-8. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. 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]
  9. Ikeda H., Kawasaki I., Gellert M. Mechanism of illegitimate recombination: common sites for recombination and cleavage mediated by E. coli DNA gyrase. Mol Gen Genet. 1984;196(3):546–549. doi: 10.1007/BF00436208. [DOI] [PubMed] [Google Scholar]
  10. Kamio Y., Tabuchi A., Itoh Y., Katagiri H., Terawaki Y. Complete nucleotide sequence of mini-Rts1 and its copy mutant. J Bacteriol. 1984 Apr;158(1):307–312. doi: 10.1128/jb.158.1.307-312.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Maki S., Miki T., Horiuchi T. DNA sequence of an amber replication mutant indicates that a 29 kd protein is the product of the F plasmid replication gene. Mol Gen Genet. 1984;194(1-2):337–339. doi: 10.1007/BF00383537. [DOI] [PubMed] [Google Scholar]
  12. Messing J., Vieira J. A new pair of M13 vectors for selecting either DNA strand of double-digest restriction fragments. Gene. 1982 Oct;19(3):269–276. doi: 10.1016/0378-1119(82)90016-6. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Murotsu T., Tsutsui H., Matsubara K. Identification of the minimal essential region for the replication origin of miniF plasmid. Mol Gen Genet. 1984;196(2):373–378. doi: 10.1007/BF00328075. [DOI] [PubMed] [Google Scholar]
  15. Scott J. R. Regulation of plasmid replication. Microbiol Rev. 1984 Mar;48(1):1–23. doi: 10.1016/b978-0-12-048850-6.50006-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Som T., Tomizawa J. Origin of replication of Escherichia coli plasmid RSF 1030. Mol Gen Genet. 1982;187(3):375–383. doi: 10.1007/BF00332615. [DOI] [PubMed] [Google Scholar]
  17. Sternberg N., Austin S. Isolation and characterization of P1 minireplicons, lambda-P1:5R and lambda-P1:5L. J Bacteriol. 1983 Feb;153(2):800–812. doi: 10.1128/jb.153.2.800-812.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Søgaard-Andersen L., Rokeach L. A., Molin S. Regulated expression of a gene important for replication of plasmid F in E. coli. EMBO J. 1984 Feb;3(2):257–262. doi: 10.1002/j.1460-2075.1984.tb01794.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Tsutsui H., Fujiyama A., Murotsu T., Matsubara K. Role of nine repeating sequences of the mini-F genome for expression of F-specific incompatibility phenotype and copy number control. J Bacteriol. 1983 Jul;155(1):337–344. doi: 10.1128/jb.155.1.337-344.1983. [DOI] [PMC free article] [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