1887

Abstract

The streptococcal plasmid pMV158 encodes a gene cassette involved in its mobilization by large conjugative plasmids. Two elements compose this region: i) the gene, encoding the MobM protein that initiates transfer, and ii) the origin of transfer, , which is the target of MobM. , MobM protein introduces a specific nick within the pMV158- region on supercoiled pMV158 DNA. This paper reports the uncoupling of the and the gene, the latter being placed under the control of an inducible promoter. Upon induction, the vector containing pMV158- was transferred in matings at a moderate frequency whereas, , purified MobM protein efficiently cleaved the vector harbouring the pMV158-. Transfer of this vector, as well as transfer of pMV158 in , required the presence of either the IncW R388 or the IncP RP4 plasmids as auxiliary plasmids. Dissection of the functions encoded by RP4 showed that the and genes were essential for pMV158 mobilization.

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2000-09-01
2024-04-23
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References

  1. Balzer D., Pansegrau W., Lamka E. 1994; Essential motifs of relaxase (TraI) and TraG proteins involved in conjugative transfer of plasmid RP4. J Bacteriol 176:4285–4295
    [Google Scholar]
  2. Bates S., Roscoe R. A., Althorpe N. J., Brammar W. J., Wilkins B. M. 1999; Expression of leading region genes on IncI1 plasmid ColIb-P9: genetic evidence for single-stranded DNA transcription. Microbiology 145:2655–2662
    [Google Scholar]
  3. Bertram J., Strätz M., Dürre P. 1991; Natural transfer of conjugative transposon Tn916 between gram-positive and gram-negative bacteria. J Bacteriol 173:443–448
    [Google Scholar]
  4. Bolland S., Llosa M., Avila P., de la Cruz F. 1990; General organization of the conjugal transfer genes of the IncW plasmid R388 and interactions between R388 and IncN and IncP plasmids. J Bacteriol 172:5795–5802
    [Google Scholar]
  5. Burdett V. 1980; Identification of tetracycline-resistant R-plasmids in Streptococcus agalactiae (group B). Antimicrob Agents Chemother 18:753–760 [CrossRef]
    [Google Scholar]
  6. Campbell J. L., Richardson C. C., Studier F. W. 1978; Genetic recombination and complementation between bacteriophage T7 and cloned fragments of T7 DNA. Proc Natl Acad Sci USA 75:2276–2280 [CrossRef]
    [Google Scholar]
  7. Crellin P. K., Rood J. I. 1998; Tn4451 from Clostridium perfringens is a mobilizable transposon that encodes the functional Mob protein, TnpZ. Mol Microbiol 27:631–642 [CrossRef]
    [Google Scholar]
  8. Datta N., Hedges R. W. 1972; Trimethoprim resistance conferred by W plasmids in Enterobacteriaceae. J Gen Microbiol 72:349–355 [CrossRef]
    [Google Scholar]
  9. Farı́as M. E., Grohmann E., Espinosa M. 1999; Expression of the mobM gene of the streptococcal plasmid pMV158 in Lactococcus lactis subsp. lactis. FEMS Microbiol Lett 176:403–410 [CrossRef]
    [Google Scholar]
  10. Grohmann E., Zechner E. L., Espinosa M. 1997; Determination of specific DNA strand discontinuities with nucleotide resolution in exponentially growing bacteria harbouring rolling circle-replicating plasmids. FEMS Microbiol Lett 152:363–369 [CrossRef]
    [Google Scholar]
  11. Grohmann E., Guzmán L. M., Espinosa M. 1999; Mobilisation of the streptococcal plasmid pMV158: interactions of MobM protein with its cognate oriT DNA region. Mol Gen Genet 261:707–715 [CrossRef]
    [Google Scholar]
  12. Guzmán L., Espinosa M. 1997; The mobilization protein, MobM, of the streptococcal plasmid pMV158 specifically cleaves supercoiled DNA at the plasmid oriT. J Mol Biol 267:688–702
    [Google Scholar]
  13. Lacks S. A., López P., Greenberg B., Espinosa M. 1986; Identification and analysis of genes for tetracycline resistance and replication functions in the broad-host-range plasmid pLS1. J Mol Biol 192:753–765 [CrossRef]
    [Google Scholar]
  14. Lanka E., Wilkins B. M. 1995; DNA processing reactions in bacterial conjugation. Annu Rev Biochem 64:141–169 [CrossRef]
    [Google Scholar]
  15. LeBlanc D. J., Lee L. N., Abu-Al-Jaibat A. 1992; Molecular, genetic, and functional analysis of the basic replicon of pVA380-1, a plasmid of oral streptococcal origin. Plasmid 28:130–145 [CrossRef]
    [Google Scholar]
  16. van der Lelie D., Wosten H. A. B., Bron S., Oskam L., Venema G. 1990; Conjugal mobilization of streptococcal plasmid pMV158 between strains of Lactococcus lactis subsp. lactis. J Bacteriol 172:47–52
    [Google Scholar]
  17. Llosa M., Grandoso G., de la Cruz F. 1995; Nicking activity of TrwC directed against the origin of transfer of the IncW plasmid R388. J Mol Biol 246:54–62 [CrossRef]
    [Google Scholar]
  18. Masai H., Arai K. 1997; Frpo: a novel single-stranded DNA promoter for transcription and for primer RNA synthesis of DNA replication. Cell 89:897–907 [CrossRef]
    [Google Scholar]
  19. Moscoso M., del Solar G., Espinosa M. 1995; In vitro recognition of the replication origin of pLS1 and of plasmids of the pLS1 family by the RepB initiator protein. J Bacteriol 177:7041–7049
    [Google Scholar]
  20. Murphy C. G., Mallamy M. H. 1993; Characterization of a ‘mobilization cassette’ in transposon Tn4399 from Bacteroides fragilis. J Bacteriol 175:5814–5823
    [Google Scholar]
  21. Pansegrau W., Lanka E. 1996; Enzymology of DNA strand transfer by conjugative mechanisms. Prog Nucleic Acid Res Mol Biol 54:197–251
    [Google Scholar]
  22. Pansegrau W., Lanka E., Barth P. T.7 other authors 1994; Complete nucleotide sequence of Birmingham IncPα plasmids: compilation and comparative analysis. J Mol Biol 239:623–662 [CrossRef]
    [Google Scholar]
  23. Priebe S. D., Lacks S. A. 1989; Region of the streptococcal plasmid pMV158 required for conjugative mobilization. J Bacteriol 171:4778–4784
    [Google Scholar]
  24. Sambrook J., Fritsch E. F., Maniatis T. 1989 Molecular Cloning: a Laboratory Manual, 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  25. Smith C. J., Tribble G. D., Bayley D. P. 1998; Genetic elements of Bacteroides species: a moving story. Plasmid 40:12–29 [CrossRef]
    [Google Scholar]
  26. Smith M. D., Shoemaker N. B., Burdett V., Guild W. R. 1980; Transfer of plasmids by conjugation in Streptococcus pneumoniae. Plasmid 3:70–79 [CrossRef]
    [Google Scholar]
  27. del Solar G., Dı́az R., Espinosa M. 1987; Replication of the streptococcal plasmid pMV158 and derivatives in cell-free extracts of Escherichia coli. Mol Gen Genet 206:428–435 [CrossRef]
    [Google Scholar]
  28. del Solar G., Giraldo R., Ruiz-Echevarrı́a M. J., Espinosa M., Dı́az-Orejas R. 1998; Replication and control of circular bacterial plasmids. Microbiol Mol Biol Rev 62:434–464
    [Google Scholar]
  29. Trieu-Cuot P., Arthur M., Courvalin P. 1987; Plasmid transfer by conjugation from Escherichia coli to gram-positive bacteria. FEMS Microbiol Lett 48:289–294 [CrossRef]
    [Google Scholar]
  30. Wang A., Macrina F. L. 1995; Characterization of six linked open reading frames necessary for pIP501-mediated conjugation. Plasmid 34:206–210 [CrossRef]
    [Google Scholar]
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