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. 1990 Apr;9(4):991–998. doi: 10.1002/j.1460-2075.1990.tb08201.x

Specificity switching of the P1 plasmid centromere-like site.

M A Davis 1, K A Martin 1, S J Austin 1
PMCID: PMC551768  PMID: 2182325

Abstract

The P1 plasmid partition site acts like a centromere, promoting accurate segregation of copies to daughter cells. A 34 bp segment is essential for partition and binds the plasmid ParB protein. Additional sequences act as specificity elements that direct the choice of copies for partition. They include a second ParB binding site and a site for the host integration host factor protein. Sites lacking one or more of these additional elements are switched to a different specificity. Defined mutants were scored for partition specificity and protein binding. The results suggest that the wild-type site is folded in a specific DNA-protein complex. Disruption of the complex leads to an open configuration which, while still active in partition, has altered recognition specificity.

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Selected References

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

  1. Abeles A. L., Friedman S. A., Austin S. J. Partition of unit-copy miniplasmids to daughter cells. III. The DNA sequence and functional organization of the P1 partition region. J Mol Biol. 1985 Sep 20;185(2):261–272. doi: 10.1016/0022-2836(85)90402-4. [DOI] [PubMed] [Google Scholar]
  2. Abeles A. L. P1 plasmid replication. Purification and DNA-binding activity of the replication protein RepA. J Biol Chem. 1986 Mar 15;261(8):3548–3555. [PubMed] [Google Scholar]
  3. Austin S. J. Plasmid partition. Plasmid. 1988 Jul;20(1):1–9. doi: 10.1016/0147-619x(88)90001-7. [DOI] [PubMed] [Google Scholar]
  4. Austin S., Abeles A. Partition of unit-copy miniplasmids to daughter cells. I. P1 and F miniplasmids contain discrete, interchangeable sequences sufficient to promote equipartition. J Mol Biol. 1983 Sep 15;169(2):353–372. doi: 10.1016/s0022-2836(83)80055-2. [DOI] [PubMed] [Google Scholar]
  5. Austin S., Ziese M., Sternberg N. A novel role for site-specific recombination in maintenance of bacterial replicons. Cell. 1981 Sep;25(3):729–736. doi: 10.1016/0092-8674(81)90180-x. [DOI] [PubMed] [Google Scholar]
  6. Davis M. A., Austin S. J. Recognition of the P1 plasmid centromere analog involves binding of the ParB protein and is modified by a specific host factor. EMBO J. 1988 Jun;7(6):1881–1888. doi: 10.1002/j.1460-2075.1988.tb03021.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dennert G., Henning U. Tyrosine-incorporating amber suppressors in Escherichia coli K12. J Mol Biol. 1968 Apr 14;33(1):327–329. doi: 10.1016/0022-2836(68)90300-8. [DOI] [PubMed] [Google Scholar]
  8. Friedman D. I. Integration host factor: a protein for all reasons. Cell. 1988 Nov 18;55(4):545–554. doi: 10.1016/0092-8674(88)90213-9. [DOI] [PubMed] [Google Scholar]
  9. Funnell B. E. Mini-P1 plasmid partitioning: excess ParB protein destabilizes plasmids containing the centromere parS. J Bacteriol. 1988 Feb;170(2):954–960. doi: 10.1128/jb.170.2.954-960.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Funnell B. E. Participation of Escherichia coli integration host factor in the P1 plasmid partition system. Proc Natl Acad Sci U S A. 1988 Sep;85(18):6657–6661. doi: 10.1073/pnas.85.18.6657. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Galas D. J., Schmitz A. DNAse footprinting: a simple method for the detection of protein-DNA binding specificity. Nucleic Acids Res. 1978 Sep;5(9):3157–3170. doi: 10.1093/nar/5.9.3157. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Gamas P., Chandler M. G., Prentki P., Galas D. J. Escherichia coli integration host factor binds specifically to the ends of the insertion sequence IS1 and to its major insertion hot-spot in pBR322. J Mol Biol. 1987 May 20;195(2):261–272. doi: 10.1016/0022-2836(87)90648-6. [DOI] [PubMed] [Google Scholar]
  13. Gardner J. F., Nash H. A. Role of Escherichia coli IHF protein in lambda site-specific recombination. A mutational analysis of binding sites. J Mol Biol. 1986 Sep 20;191(2):181–189. doi: 10.1016/0022-2836(86)90255-x. [DOI] [PubMed] [Google Scholar]
  14. Gottesman M. E., Yarmolinsky M. B. Integration-negative mutants of bacteriophage lambda. J Mol Biol. 1968 Feb 14;31(3):487–505. doi: 10.1016/0022-2836(68)90423-3. [DOI] [PubMed] [Google Scholar]
  15. Martin K. A., Friedman S. A., Austin S. J. Partition site of the P1 plasmid. Proc Natl Acad Sci U S A. 1987 Dec;84(23):8544–8547. doi: 10.1073/pnas.84.23.8544. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. 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]
  17. Nash H. A., Robertson C. A., Flamm E., Weisberg R. A., Miller H. I. Overproduction of Escherichia coli integration host factor, a protein with nonidentical subunits. J Bacteriol. 1987 Sep;169(9):4124–4127. doi: 10.1128/jb.169.9.4124-4127.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Rao R. N. Construction and properties of plasmid pKC30, a pBR322 derivative containing the pL-N region of phage lambda. Gene. 1984 Nov;31(1-3):247–250. doi: 10.1016/0378-1119(84)90216-6. [DOI] [PubMed] [Google Scholar]
  19. Rosner J. L. Formation, induction, and curing of bacteriophage P1 lysogens. Virology. 1972 Jun;48(3):679–689. doi: 10.1016/0042-6822(72)90152-3. [DOI] [PubMed] [Google Scholar]
  20. Yang C. C., Nash H. A. The interaction of E. coli IHF protein with its specific binding sites. Cell. 1989 Jun 2;57(5):869–880. doi: 10.1016/0092-8674(89)90801-5. [DOI] [PubMed] [Google Scholar]

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