Silencing and activating type IV secretion genes of the F-like conjugative resistance plasmid R1 Free

Abstract

Expression of DNA transfer () genes of F-type conjugative plasmids is required for the assembly of a functional type IV secretion machinery and subsequent plasmid DNA transfer from donor to recipient cells. Transcription of genes depends on the activation of a single promoter, designated P, by the plasmid encoded TraJ protein. We here determine plasmid specificity of TraJ proteins from various subgroups of F-like plasmids and find that plasmid R1 conjugation and P promoter activation can be achieved only by its cognate activator and by TraJ of the plasmid pSLT and not by F or R100 TraJ proteins. In addition, we characterize the P promoter of plasmid R1. We show that TraJ binds to P DNA and that H-NS acts as a silencer of the P promoter. In the natural plasmid context, H-NS silences transfer gene expression and horizontal plasmid DNA transfer. In contrast to what was found for the F plasmid, lack of H-NS did not abolish the requirement for ArcA and TraJ to reach full gene expression and DNA transfer activity. We propose that, besides a passive de-silencing activity, both ArcA and TraJ play a direct role in synergistically stimulating operon transcription and subsequent DNA transfer.

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2013-12-01
2024-03-29
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References

  1. Arthur D. C., Ghetu A. F., Gubbins M. J., Edwards R. A., Frost L. S., Glover J. N.( 2003). FinO is an RNA chaperone that facilitates sense–antisense RNA interactions. EMBO J 22:6346–6355 [View Article][PubMed]
    [Google Scholar]
  2. Barnard A., Wolfe A., Busby S.( 2004). Regulation at complex bacterial promoters: how bacteria use different promoter organizations to produce different regulatory outcomes. Curr Opin Microbiol 7:102–108 [View Article][PubMed]
    [Google Scholar]
  3. Barth M., Marschall C., Muffler A., Fischer D., Hengge-Aronis R.( 1995). Role for the histone-like protein H-NS in growth phase-dependent and osmotic regulation of σS and many σS-dependent genes in Escherichia coli. J Bacteriol 177:3455–3464[PubMed]
    [Google Scholar]
  4. Borukhov S., Nudler E.( 2008). RNA polymerase: the vehicle of transcription. Trends Microbiol 16:126–134 [View Article][PubMed]
    [Google Scholar]
  5. Browning D. F., Busby S. J.( 2004). The regulation of bacterial transcription initiation. Nat Rev Microbiol 2:57–65 [View Article][PubMed]
    [Google Scholar]
  6. Camacho E. M., Casadesús J.( 2002). Conjugal transfer of the virulence plasmid of Salmonella enterica is regulated by the leucine-responsive regulatory protein and DNA adenine methylation. Mol Microbiol 44:1589–1598 [View Article][PubMed]
    [Google Scholar]
  7. Cheah K. C., Skurray R.( 1986). The F plasmid carries an IS3 insertion within finO.. J Gen Microbiol 132:3269–3275[PubMed]
    [Google Scholar]
  8. de la Cruz F., Frost L. S., Meyer R. J., Zechner E. L.( 2010). Conjugative DNA metabolism in Gram-negative bacteria. FEMS Microbiol Rev 34:18–40 [View Article][PubMed]
    [Google Scholar]
  9. Dove S. L., Darst S. A., Hochschild A.( 2003). Region 4 of σ as a target for transcription regulation. Mol Microbiol 48:863–874 [View Article][PubMed]
    [Google Scholar]
  10. Feklistov A., Darst S. A.( 2011). Structural basis for promoter −10 element recognition by the bacterial RNA polymerase σ subunit. Cell 147:1257–1269 [View Article][PubMed]
    [Google Scholar]
  11. Frost L. S., Koraimann G.( 2010). Regulation of bacterial conjugation: balancing opportunity with adversity. Future Microbiol 5:1057–1071 [View Article][PubMed]
    [Google Scholar]
  12. Frost L. S., Manchak J.( 1998). F phenocopies: characterization of expression of the F transfer region in stationary phase. Microbiology 144:2579–2587 [View Article][PubMed]
    [Google Scholar]
  13. García-Quintanilla M., Ramos-Morales F., Casadesús J.( 2008). Conjugal transfer of the Salmonella enterica virulence plasmid in the mouse intestine. J Bacteriol 190:1922–1927 [View Article][PubMed]
    [Google Scholar]
  14. Grainger D. C., Overton T. W., Reppas N., Wade J. T., Tamai E., Hobman J. L., Constantinidou C., Struhl K., Church G., Busby S. J. W.( 2004). Genomic studies with Escherichia coli MelR protein: applications of chromatin immunoprecipitation and microarrays. J Bacteriol 186:6938–6943 [View Article][PubMed]
    [Google Scholar]
  15. Hinton D. M.( 2005). Molecular gymnastics: distortion of an RNA polymerase σ factor. Trends Microbiol 13:140–143 [View Article][PubMed]
    [Google Scholar]
  16. Koraimann G.( 2004). Bacterial conjugation: cell-to-cell contact dependent horizontal gene spread. Microbial Evolution Gene Establishment, Survival and Exchange111–124 Miller R. V., Day M. J. Washington, DC: American Society for Microbiology;
    [Google Scholar]
  17. Koraimann G., Högenauer G.( 1989). A stable core region of the tra operon mRNA of plasmid R1-19. Nucleic Acids Res 17:1283–1298 [View Article][PubMed]
    [Google Scholar]
  18. Koraimann G., Teferle K., Markolin G., Woger W., Högenauer G.( 1996). The FinOP repressor system of plasmid R1: analysis of the antisense RNA control of traJ expression and conjugative DNA transfer. Mol Microbiol 21:811–821 [View Article][PubMed]
    [Google Scholar]
  19. Lawley T., Wilkins B. M., Frost L.( 2004). Conjugation in Gram-negative bacteria. Plasmid Biology203–226 Phillips G., Funnell B. Washington, DC: American Society for Microbiology;
    [Google Scholar]
  20. Lessl M., Balzer D., Lurz R., Waters V. L., Guiney D. G., Lanka E.( 1992). Dissection of IncP conjugative plasmid transfer: definition of the transfer region Tra2 by mobilization of the Tra1 region in trans.. J Bacteriol 174:2493–2500[PubMed]
    [Google Scholar]
  21. Liu X., De Wulf P.( 2004). Probing the ArcA–P modulon of Escherichia coli by whole genome transcriptional analysis and sequence recognition profiling. J Biol Chem 279:12588–12597 [View Article][PubMed]
    [Google Scholar]
  22. Livak K. J., Schmittgen T. D.( 2001). Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 25:402–408 [View Article][PubMed]
    [Google Scholar]
  23. Miller J. H.( 1972). Experiments in Molecular Genetics Cold Spring Harbor, New York: Cold Spring Harbor Laboratory Press;
    [Google Scholar]
  24. Nelson W. C., Morton B. S., Lahue E. E., Matson S. W.( 1993). Characterization of the Escherichia coli F factor traY gene product and its binding sites. J Bacteriol 175:2221–2228[PubMed]
    [Google Scholar]
  25. Nordström K.( 2006). Plasmid R1 – replication and its control. Plasmid 55:1–26 [View Article][PubMed]
    [Google Scholar]
  26. Pagni M., Ioannidis V., Cerutti L., Zahn-Zabal M., Jongeneel C. V., Hau J., Martin O., Kuznetsov D., Falquet L.( 2007). MyHits: improvements to an interactive resource for analyzing protein sequences. Nucleic Acids Res 35:Web Server issueW433–W437 [View Article][PubMed]
    [Google Scholar]
  27. Pande S., Makela A., Dove S. L., Nickels B. E., Hochschild A., Hinton D. M.( 2002). The bacteriophage T4 transcription activator MotA interacts with the far-C-terminal region of the σ70 subunit of Escherichia coli RNA polymerase. J Bacteriol 184:3957–3964 [View Article][PubMed]
    [Google Scholar]
  28. Reisner A., Höller B. M., Molin S., Zechner E. L.( 2006). Synergistic effects in mixed Escherichia coli biofilms: conjugative plasmid transfer drives biofilm expansion. J Bacteriol 188:3582–3588 [View Article][PubMed]
    [Google Scholar]
  29. Rodriguez-Maillard J. M., Arutyunov D., Frost L. S.( 2010). The F plasmid transfer activator TraJ is a dimeric helix-turn-helix DNA-binding protein. FEMS Microbiol Lett 310:112–119 [View Article][PubMed]
    [Google Scholar]
  30. Salmon K. A., Hung S. P., Steffen N. R., Krupp R., Baldi P., Hatfield G. W., Gunsalus R. P.( 2005). Global gene expression profiling in Escherichia coli K12: effects of oxygen availability and ArcA. J Biol Chem 280:15084–15096 [View Article][PubMed]
    [Google Scholar]
  31. Sambrook J., Fritsch E. F., Maniatis T.( 1989). Molecular Cloning: a Laboratory Manual, 2nd edn. Cold Spring Harbor, New York: Cold Spring Harbor Laboratory Press;
    [Google Scholar]
  32. Serna A., Espinosa E., Camacho E. M., Casadesús J.( 2010). Regulation of bacterial conjugation in microaerobiosis by host-encoded functions ArcAB and SdhABCD. Genetics 184:947–958 [View Article][PubMed]
    [Google Scholar]
  33. Silverman P. M., Wickersham E., Harris R.( 1991). Regulation of the F plasmid traY promoter in Escherichia coli by host and plasmid factors. J Mol Biol 218:119–128 [View Article][PubMed]
    [Google Scholar]
  34. Starcic M., Zgur-Bertok D., Jordi B. J., Wösten M. M., Gaastra W., van Putten J. P.( 2003). The cyclic AMP–cyclic AMP receptor protein complex regulates activity of the traJ promoter of the Escherichia coli conjugative plasmid pRK100. J Bacteriol 185:1616–1623 [View Article][PubMed]
    [Google Scholar]
  35. Strohmaier H., Noiges R., Kotschan S., Sawers G., Högenauer G., Zechner E. L., Koraimann G.( 1998). Signal transduction and bacterial conjugation: characterization of the role of ArcA in regulating conjugative transfer of the resistance plasmid R1. J Mol Biol 277:309–316 [View Article][PubMed]
    [Google Scholar]
  36. Taki K., Abo T., Ohtsubo E.( 1998). Regulatory mechanisms in expression of the traY–I operon of sex factor plasmid R100: involvement of traJ and traY gene products. Genes Cells 3:331–345 [View Article][PubMed]
    [Google Scholar]
  37. van Hijum S. A., Medema M. H., Kuipers O. P.( 2009). Mechanisms and evolution of control logic in prokaryotic transcriptional regulation. Microbiol Mol Biol Rev 73:481–509 [View Article][PubMed]
    [Google Scholar]
  38. Wagner M. A., Zahrl D., Rieser G., Koraimann G.( 2009). Growth phase- and cell division-dependent activation and inactivation of the σ32 regulon in Escherichia coli. J Bacteriol 191:1695–1702 [View Article][PubMed]
    [Google Scholar]
  39. Will W. R., Frost L. S.( 2006). Characterization of the opposing roles of H-NS and TraJ in transcriptional regulation of the F-plasmid tra operon. J Bacteriol 188:507–514 [View Article][PubMed]
    [Google Scholar]
  40. Will W. R., Lu J., Frost L. S.( 2004). The role of H-NS in silencing F transfer gene expression during entry into stationary phase. Mol Microbiol 54:769–782 [View Article][PubMed]
    [Google Scholar]
  41. Wong J. J. W., Lu J., Glover J. N. M.( 2012). Relaxosome function and conjugation regulation in F-like plasmids – a structural biology perspective. Mol Microbiol 85:602–617 [View Article][PubMed]
    [Google Scholar]
  42. Zahrl D., Wagner M., Bischof K., Koraimann G.( 2006). Expression and assembly of a functional type IV secretion system elicit extracytoplasmic and cytoplasmic stress responses in Escherichia coli.. J Bacteriol 188:6611–6621 [View Article][PubMed]
    [Google Scholar]
  43. Zahrl D., Wagner A., Tscherner M., Koraimann G.( 2007). GroEL plays a central role in stress-induced negative regulation of bacterial conjugation by promoting proteolytic degradation of the activator protein TraJ. J Bacteriol 189:5885–5894 [View Article][PubMed]
    [Google Scholar]
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