Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1987 Jun;169(6):2601–2610. doi: 10.1128/jb.169.6.2601-2610.1987

A site-specific recombination function in Staphylococcus aureus plasmids.

M L Gennaro, J Kornblum, R P Novick
PMCID: PMC212133  PMID: 3584064

Abstract

All known small staphylococcal plasmids possess one or two recombination sites at which site-specific cointegrate formation occurs. One of these sites, RSA, is present on two small multicopy plasmids, pT181 and pE194; it consists of 24 base pairs of identity in the two plasmids, the "core," flanked by some 50 base pairs of decreasing homology. Here we show that recombination at RSA is recA independent and is mediated by a plasmid-encoded, trans-acting protein, Pre (plasmid recombination). Pre-mediated recombination is site specific in that it occurs within the core sequence of RSA in a recA1 host. Recombination also occurs between two intramolecular RSA sites. Unlike site-specific recombination systems encoded by other plasmids, Pre-RSA is not involved in plasmid maintenance.

Full text

PDF
2601

Images in this article

Selected References

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

  1. Arthur A., Sherratt D. Dissection of the transposition process: a transposon-encoded site-specific recombination system. Mol Gen Genet. 1979 Oct 1;175(3):267–274. doi: 10.1007/BF00397226. [DOI] [PubMed] [Google Scholar]
  2. 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]
  3. Bhaduri S., Demchick P. H. Simple and rapid method for disruption of bacteria for protein studies. Appl Environ Microbiol. 1983 Oct;46(4):941–943. doi: 10.1128/aem.46.4.941-943.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Carleton S., Projan S. J., Highlander S. K., Moghazeh S. M., Novick R. P. Control of pT181 replication II. Mutational analysis. EMBO J. 1984 Oct;3(10):2407–2414. doi: 10.1002/j.1460-2075.1984.tb02147.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chang S., Cohen S. N. High frequency transformation of Bacillus subtilis protoplasts by plasmid DNA. Mol Gen Genet. 1979 Jan 5;168(1):111–115. doi: 10.1007/BF00267940. [DOI] [PubMed] [Google Scholar]
  6. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  7. Fisher R. F., Das A., Kolter R., Winkler M. E., Yanofsky C. Analysis of the requirements for transcription pausing in the tryptophan operon. J Mol Biol. 1985 Apr 5;182(3):397–409. doi: 10.1016/0022-2836(85)90199-8. [DOI] [PubMed] [Google Scholar]
  8. Glisin V., Crkvenjakov R., Byus C. Ribonucleic acid isolated by cesium chloride centrifugation. Biochemistry. 1974 Jun 4;13(12):2633–2637. doi: 10.1021/bi00709a025. [DOI] [PubMed] [Google Scholar]
  9. Hakkaart M. J., van den Elzen P. J., Veltkamp E., Nijkamp H. J. Maintenance of multicopy plasmid Clo DF13 in E. coli cells: evidence for site-specific recombination at parB. Cell. 1984 Jan;36(1):203–209. doi: 10.1016/0092-8674(84)90090-4. [DOI] [PubMed] [Google Scholar]
  10. Horinouchi S., Weisblum B. Nucleotide sequence and functional map of pC194, a plasmid that specifies inducible chloramphenicol resistance. J Bacteriol. 1982 May;150(2):815–825. doi: 10.1128/jb.150.2.815-825.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Horinouchi S., Weisblum B. Nucleotide sequence and functional map of pE194, a plasmid that specifies inducible resistance to macrolide, lincosamide, and streptogramin type B antibodies. J Bacteriol. 1982 May;150(2):804–814. doi: 10.1128/jb.150.2.804-814.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Iida S., Meyer J., Kennedy K. E., Arber W. A site-specific, conservative recombination system carried by bacteriophage P1. Mapping the recombinase gene cin and the cross-over sites cix for the inversion of the C segment. EMBO J. 1982;1(11):1445–1453. doi: 10.1002/j.1460-2075.1982.tb01336.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Iordănescu S. Recombinant plasmid obtained from two different, compatible staphylococcal plasmids. J Bacteriol. 1975 Nov;124(2):597–601. doi: 10.1128/jb.124.2.597-601.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Iordănescu S., Surdeanu M. Relationships between autonomous and integrated forms of tetracycline resistance plasmid in Staphylococcus aureus. Plasmid. 1979 Apr;2(2):216–224. doi: 10.1016/0147-619x(79)90040-4. [DOI] [PubMed] [Google Scholar]
  15. Iordănescu S. Three distinct plasmids originating in the same Staphylococcus aureus strain. Arch Roum Pathol Exp Microbiol. 1976 Jan-Jun;35(1-2):111–118. [PubMed] [Google Scholar]
  16. Kamp D., Chow L. T., Broker T. R., Kwoh D., Zipser D., Kahmann R. Site-specific recombination in phage mu. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 2):1159–1167. doi: 10.1101/sqb.1979.043.01.131. [DOI] [PubMed] [Google Scholar]
  17. Khan S. A., Novick R. P. Complete nucleotide sequence of pT181, a tetracycline-resistance plasmid from Staphylococcus aureus. Plasmid. 1983 Nov;10(3):251–259. doi: 10.1016/0147-619x(83)90039-2. [DOI] [PubMed] [Google Scholar]
  18. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  19. Machida Y., Machida C., Ohtsubo H., Ohtsubo E. Factors determining frequency of plasmid cointegration mediated by insertion sequence IS1. Proc Natl Acad Sci U S A. 1982 Jan;79(2):277–281. doi: 10.1073/pnas.79.2.277. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  21. McKenzie T., Hoshino T., Tanaka T., Sueoka N. Correction. A revision of the nucleotide sequence and functional map of pUB110. Plasmid. 1987 Jan;17(1):83–85. doi: 10.1016/0147-619x(87)90015-1. [DOI] [PubMed] [Google Scholar]
  22. Novick R. P., Adler G. K., Majumder S., Khan S. A., Carleton S., Rosenblum W. D., Iordanescu S. Coding sequence for the pT181 repC product: a plasmid-coded protein uniquely required for replication. Proc Natl Acad Sci U S A. 1982 Jul;79(13):4108–4112. doi: 10.1073/pnas.79.13.4108. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Novick R. P., Adler G. K., Projan S. J., Carleton S., Highlander S. K., Gruss A., Khan S. A., Iordanescu S. Control of pT181 replication I. The pT181 copy control function acts by inhibiting the synthesis of a replication protein. EMBO J. 1984 Oct;3(10):2399–2405. doi: 10.1002/j.1460-2075.1984.tb02146.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Novick R. P., Iordanescu S., Surdeanu M., Edelman I. Transduction-related cointegrate formation between Staphylococcal plasmids: a new type of site-specific recombination. Plasmid. 1981 Sep;6(2):159–172. doi: 10.1016/0147-619x(81)90064-0. [DOI] [PubMed] [Google Scholar]
  25. Novick R. P., Murphy E., Gryczan T. J., Baron E., Edelman I. Penicillinase plasmids of Staphylococcus aureus: restriction-deletion maps. Plasmid. 1979 Jan;2(1):109–129. doi: 10.1016/0147-619x(79)90010-6. [DOI] [PubMed] [Google Scholar]
  26. Novick R. P., Projan S. J., Rosenblum W., Edelman I. Staphylococcal plasmid cointegrates are formed by host- and phage-mediated general rec systems that act on short regions of homology. Mol Gen Genet. 1984;195(1-2):374–377. doi: 10.1007/BF00332777. [DOI] [PubMed] [Google Scholar]
  27. Novick R. Properties of a cryptic high-frequency transducing phage in Staphylococcus aureus. Virology. 1967 Sep;33(1):155–166. doi: 10.1016/0042-6822(67)90105-5. [DOI] [PubMed] [Google Scholar]
  28. Plasterk R. H., Van de Putte P. Genetic switches by DNA inversions in prokaryotes. Biochim Biophys Acta. 1984 Jun 16;782(2):111–119. doi: 10.1016/0167-4781(84)90013-7. [DOI] [PubMed] [Google Scholar]
  29. Polak J., Novick R. P. Closely related plasmids from Staphylococcus aureus and soil bacilli. Plasmid. 1982 Mar;7(2):152–162. doi: 10.1016/0147-619x(82)90074-9. [DOI] [PubMed] [Google Scholar]
  30. Projan S. J., Carleton S., Novick R. P. Determination of plasmid copy number by fluorescence densitometry. Plasmid. 1983 Mar;9(2):182–190. doi: 10.1016/0147-619x(83)90019-7. [DOI] [PubMed] [Google Scholar]
  31. Reed R. R. Resolution of cointegrates between transposons gamma delta and Tn3 defines the recombination site. Proc Natl Acad Sci U S A. 1981 Jun;78(6):3428–3432. doi: 10.1073/pnas.78.6.3428. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  33. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Shivakumar A. G., Gryczan T. J., Kozlov Y. I., Dubnau D. Organization of the pE194 genome. Mol Gen Genet. 1980;179(2):241–252. doi: 10.1007/BF00425450. [DOI] [PubMed] [Google Scholar]
  35. Sternberg N., Hamilton D., Austin S., Yarmolinsky M., Hoess R. Site-specific recombination and its role in the life cycle of bacteriophage P1. Cold Spring Harb Symp Quant Biol. 1981;45(Pt 1):297–309. doi: 10.1101/sqb.1981.045.01.042. [DOI] [PubMed] [Google Scholar]
  36. Studier F. W., Moffatt B. A. Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes. J Mol Biol. 1986 May 5;189(1):113–130. doi: 10.1016/0022-2836(86)90385-2. [DOI] [PubMed] [Google Scholar]
  37. Summers D. K., Sherratt D. J. Multimerization of high copy number plasmids causes instability: CoIE1 encodes a determinant essential for plasmid monomerization and stability. Cell. 1984 Apr;36(4):1097–1103. doi: 10.1016/0092-8674(84)90060-6. [DOI] [PubMed] [Google Scholar]
  38. Takahashi H., Saito H. High-frequency transduction of pBR322 by cytosine-substituted T4 bacteriophage: evidence for encapsulation and transfer of head-to-tail plasmid concatemers. Plasmid. 1982 Jul;8(1):29–35. doi: 10.1016/0147-619x(82)90038-5. [DOI] [PubMed] [Google Scholar]
  39. Wyman L., Goering R. V., Novick R. P. Genetic control of chromosomal and plasmid recombination in Staphylococcus aureus. Genetics. 1974 Apr;76(4):681–702. doi: 10.1093/genetics/76.4.681. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Wyman L., Novick R. P. Studies on plasmid replication. IV. Complementation of replication-defective mutants by an incompatibility-deficient plasmid. Mol Gen Genet. 1974;135(2):149–161. doi: 10.1007/BF00264782. [DOI] [PubMed] [Google Scholar]
  41. Zieg J., Silverman M., Hilmen M., Simon M. Recombinational switch for gene expression. Science. 1977 Apr 8;196(4286):170–172. doi: 10.1126/science.322276. [DOI] [PubMed] [Google Scholar]

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

RESOURCES