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. 1995 May;177(9):2381–2386. doi: 10.1128/jb.177.9.2381-2386.1995

Partition of P1 plasmids in Escherichia coli mukB chromosomal partition mutants.

B E Funnell 1, L Gagnier 1
PMCID: PMC176895  PMID: 7730268

Abstract

The partition system of the low-copy-number plasmid/prophage of bacteriophage P1 encodes two proteins, ParA and ParB, and contains a DNA site called parS. ParB and the Escherichia coli protein IHF bind to parS to form the partition complex, in which parS is wrapped around ParB and IHF in a precise three-dimensional conformation. Partition can be thought of as a positioning reaction; the plasmid-encoded components ensure that at least one copy of the plasmid is positioned within each new daughter cell. We have used an E. coli chromosomal partition mutant to test whether this positioning is mediated by direct plasmid-chromosomal attachment, for example, by pairing of the partition complex that forms at parS with a bacterial attachment site. The E. coli MukB protein is required for proper chromosomal positioning, so that mukB mutants generate some cells without chromosomes (anucleate cells) at each cell division. We analyzed the plasmid distribution in nucleate and anucleate mukB cells. We found that P1 plasmids are stable in mukB mutants and that they partition into both nucleate and anucleate cells. This indicates that the P1 partition complex is not used to pair plasmids with the host chromosome and that P1 plasmids must be responsible for their own proper cellular localization, presumably through host-plasmid protein-protein interactions.

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Selected References

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  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., 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]
  3. Austin S., Nordström K. Partition-mediated incompatibility of bacterial plasmids. Cell. 1990 Feb 9;60(3):351–354. doi: 10.1016/0092-8674(90)90584-2. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. Bachmann B. J. Linkage map of Escherichia coli K-12, edition 8. Microbiol Rev. 1990 Jun;54(2):130–197. doi: 10.1128/mr.54.2.130-197.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Botta G. A., Park J. T. Evidence for involvement of penicillin-binding protein 3 in murein synthesis during septation but not during cell elongation. J Bacteriol. 1981 Jan;145(1):333–340. doi: 10.1128/jb.145.1.333-340.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Calos M. P., Lebkowski J. S., Botchan M. R. High mutation frequency in DNA transfected into mammalian cells. Proc Natl Acad Sci U S A. 1983 May;80(10):3015–3019. doi: 10.1073/pnas.80.10.3015. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dassain M., Bouché J. P. The min locus, which confers topological specificity to cell division, is not involved in its coupling with nucleoid separation. J Bacteriol. 1994 Oct;176(19):6143–6145. doi: 10.1128/jb.176.19.6143-6145.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  10. Davis M. A., Martin K. A., Austin S. J. Biochemical activities of the parA partition protein of the P1 plasmid. Mol Microbiol. 1992 May;6(9):1141–1147. doi: 10.1111/j.1365-2958.1992.tb01552.x. [DOI] [PubMed] [Google Scholar]
  11. Davis M. A., Martin K. A., Austin S. J. Specificity switching of the P1 plasmid centromere-like site. EMBO J. 1990 Apr;9(4):991–998. doi: 10.1002/j.1460-2075.1990.tb08201.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Eliasson A., Bernander R., Dasgupta S., Nordström K. Direct visualization of plasmid DNA in bacterial cells. Mol Microbiol. 1992 Jan;6(2):165–170. doi: 10.1111/j.1365-2958.1992.tb01997.x. [DOI] [PubMed] [Google Scholar]
  13. Ezaki B., Ogura T., Niki H., Hiraga S. Partitioning of a mini-F plasmid into anucleate cells of the mukB null mutant. J Bacteriol. 1991 Oct;173(20):6643–6646. doi: 10.1128/jb.173.20.6643-6646.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Funnell B. E., Gagnier L. The P1 plasmid partition complex at parS. II. Analysis of ParB protein binding activity and specificity. J Biol Chem. 1993 Feb 15;268(5):3616–3624. [PubMed] [Google Scholar]
  15. 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]
  16. 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]
  17. Funnell B. E. The P1 plasmid partition complex at parS. The influence of Escherichia coli integration host factor and of substrate topology. J Biol Chem. 1991 Aug 5;266(22):14328–14337. [PubMed] [Google Scholar]
  18. Goodman S. D., Nash H. A. Functional replacement of a protein-induced bend in a DNA recombination site. Nature. 1989 Sep 21;341(6239):251–254. doi: 10.1038/341251a0. [DOI] [PubMed] [Google Scholar]
  19. Gunsalus R. P., Zurawski G., Yanofsky C. Structural and functional analysis of cloned deoxyribonucleic acid containing the trpR-thr region of the Escherichia coli chromosome. J Bacteriol. 1979 Oct;140(1):106–113. doi: 10.1128/jb.140.1.106-113.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Helsberg M., Eichenlaub R. Twelve 43-base-pair repeats map in a cis-acting region essential for partition of plasmid mini-F. J Bacteriol. 1986 Mar;165(3):1043–1045. doi: 10.1128/jb.165.3.1043-1045.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Hiraga S. Chromosome and plasmid partition in Escherichia coli. Annu Rev Biochem. 1992;61:283–306. doi: 10.1146/annurev.bi.61.070192.001435. [DOI] [PubMed] [Google Scholar]
  22. Hiraga S., Niki H., Ogura T., Ichinose C., Mori H., Ezaki B., Jaffé A. Chromosome partitioning in Escherichia coli: novel mutants producing anucleate cells. J Bacteriol. 1989 Mar;171(3):1496–1505. doi: 10.1128/jb.171.3.1496-1505.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Ikeda H., Tomizawa J. Prophage P1, and extrachromosomal replication unit. Cold Spring Harb Symp Quant Biol. 1968;33:791–798. doi: 10.1101/sqb.1968.033.01.091. [DOI] [PubMed] [Google Scholar]
  24. Kass L. R., Yarmolinsky M. B. Segregation of functional sex factor into minicells. Proc Natl Acad Sci U S A. 1970 Jul;66(3):815–822. doi: 10.1073/pnas.66.3.815. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Lehnherr H., Maguin E., Jafri S., Yarmolinsky M. B. Plasmid addiction genes of bacteriophage P1: doc, which causes cell death on curing of prophage, and phd, which prevents host death when prophage is retained. J Mol Biol. 1993 Oct 5;233(3):414–428. doi: 10.1006/jmbi.1993.1521. [DOI] [PubMed] [Google Scholar]
  26. Lin Z., Mallavia L. P. Identification of a partition region carried by the plasmid QpH1 of Coxiella burnetii. Mol Microbiol. 1994 Aug;13(3):513–523. doi: 10.1111/j.1365-2958.1994.tb00446.x. [DOI] [PubMed] [Google Scholar]
  27. Martin K. A., Davis M. A., Austin S. Fine-structure analysis of the P1 plasmid partition site. J Bacteriol. 1991 Jun;173(12):3630–3634. doi: 10.1128/jb.173.12.3630-3634.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Moitoso de Vargas L., Kim S., Landy A. DNA looping generated by DNA bending protein IHF and the two domains of lambda integrase. Science. 1989 Jun 23;244(4911):1457–1461. doi: 10.1126/science.2544029. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Mori H., Mori Y., Ichinose C., Niki H., Ogura T., Kato A., Hiraga S. Purification and characterization of SopA and SopB proteins essential for F plasmid partitioning. J Biol Chem. 1989 Sep 15;264(26):15535–15541. [PubMed] [Google Scholar]
  30. Motallebi-Veshareh M., Rouch D. A., Thomas C. M. A family of ATPases involved in active partitioning of diverse bacterial plasmids. Mol Microbiol. 1990 Sep;4(9):1455–1463. doi: 10.1111/j.1365-2958.1990.tb02056.x. [DOI] [PubMed] [Google Scholar]
  31. Niki H., Imamura R., Kitaoka M., Yamanaka K., Ogura T., Hiraga S. E.coli MukB protein involved in chromosome partition forms a homodimer with a rod-and-hinge structure having DNA binding and ATP/GTP binding activities. EMBO J. 1992 Dec;11(13):5101–5109. doi: 10.1002/j.1460-2075.1992.tb05617.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Niki H., Jaffé A., Imamura R., Ogura T., Hiraga S. The new gene mukB codes for a 177 kd protein with coiled-coil domains involved in chromosome partitioning of E. coli. EMBO J. 1991 Jan;10(1):183–193. doi: 10.1002/j.1460-2075.1991.tb07935.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Ogura T., Hiraga S. Partition mechanism of F plasmid: two plasmid gene-encoded products and a cis-acting region are involved in partition. Cell. 1983 Feb;32(2):351–360. doi: 10.1016/0092-8674(83)90454-3. [DOI] [PubMed] [Google Scholar]
  34. Richet E., Abcarian P., Nash H. A. Synapsis of attachment sites during lambda integrative recombination involves capture of a naked DNA by a protein-DNA complex. Cell. 1988 Jan 15;52(1):9–17. doi: 10.1016/0092-8674(88)90526-0. [DOI] [PubMed] [Google Scholar]
  35. Richet E., Abcarian P., Nash H. A. The interaction of recombination proteins with supercoiled DNA: defining the role of supercoiling in lambda integrative recombination. Cell. 1986 Sep 26;46(7):1011–1021. doi: 10.1016/0092-8674(86)90700-2. [DOI] [PubMed] [Google Scholar]
  36. Robertson C. A., Nash H. A. Bending of the bacteriophage lambda attachment site by Escherichia coli integration host factor. J Biol Chem. 1988 Mar 15;263(8):3554–3557. [PubMed] [Google Scholar]
  37. 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]
  38. 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]
  39. 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]
  40. Thompson J. F., Landy A. Empirical estimation of protein-induced DNA bending angles: applications to lambda site-specific recombination complexes. Nucleic Acids Res. 1988 Oct 25;16(20):9687–9705. doi: 10.1093/nar/16.20.9687. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Watanabe E., Inamoto S., Lee M. H., Kim S. U., Ogua T., Mori H., Hiraga S., Yamasaki M., Nagai K. Purification and characterization of the sopB gene product which is responsible for stable maintenance of mini-F plasmid. Mol Gen Genet. 1989 Sep;218(3):431–436. doi: 10.1007/BF00332406. [DOI] [PubMed] [Google Scholar]
  42. Watanabe E., Wachi M., Yamasaki M., Nagai K. ATPase activity of SopA, a protein essential for active partitioning of F plasmid. Mol Gen Genet. 1992 Sep;234(3):346–352. doi: 10.1007/BF00538693. [DOI] [PubMed] [Google Scholar]
  43. Williams D. R., Thomas C. M. Active partitioning of bacterial plasmids. J Gen Microbiol. 1992 Jan;138(1):1–16. doi: 10.1099/00221287-138-1-1. [DOI] [PubMed] [Google Scholar]
  44. de Boer P. A., Crossley R. E., Hand A. R., Rothfield L. I. The MinD protein is a membrane ATPase required for the correct placement of the Escherichia coli division site. EMBO J. 1991 Dec;10(13):4371–4380. doi: 10.1002/j.1460-2075.1991.tb05015.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

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