1887

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.

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.071738-0
2013-12-01
2020-11-30
Loading full text...

Full text loading...

/deliver/fulltext/micro/159/12/2481.html?itemId=/content/journal/micro/10.1099/mic.0.071738-0&mimeType=html&fmt=ahah

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 J22:6346–6355 [CrossRef][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 Microbiol7:102–108 [CrossRef][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 Bacteriol177:3455–3464[PubMed]
    [Google Scholar]
  4. Borukhov S., Nudler E..( 2008;). RNA polymerase: the vehicle of transcription. Trends Microbiol16:126–134 [CrossRef][PubMed]
    [Google Scholar]
  5. Browning D. F., Busby S. J..( 2004;). The regulation of bacterial transcription initiation. Nat Rev Microbiol2:57–65 [CrossRef][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 Microbiol44:1589–1598 [CrossRef][PubMed]
    [Google Scholar]
  7. Cheah K. C., Skurray R..( 1986;). The F plasmid carries an IS3 insertion within finO.. J Gen Microbiol132:3269–3275[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 Rev34:18–40 [CrossRef][PubMed]
    [Google Scholar]
  9. Dove S. L., Darst S. A., Hochschild A..( 2003;). Region 4 of σ as a target for transcription regulation. Mol Microbiol48:863–874 [CrossRef][PubMed]
    [Google Scholar]
  10. Feklistov A., Darst S. A..( 2011;). Structural basis for promoter −10 element recognition by the bacterial RNA polymerase σ subunit. Cell147:1257–1269 [CrossRef][PubMed]
    [Google Scholar]
  11. Frost L. S., Koraimann G..( 2010;). Regulation of bacterial conjugation: balancing opportunity with adversity. Future Microbiol5:1057–1071 [CrossRef][PubMed]
    [Google Scholar]
  12. Frost L. S., Manchak J..( 1998;). F phenocopies: characterization of expression of the F transfer region in stationary phase. Microbiology144:2579–2587 [CrossRef][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 Bacteriol190:1922–1927 [CrossRef][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 Bacteriol186:6938–6943 [CrossRef][PubMed]
    [Google Scholar]
  15. Hinton D. M..( 2005;). Molecular gymnastics: distortion of an RNA polymerase σ factor. Trends Microbiol13:140–143 [CrossRef][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 Res17:1283–1298 [CrossRef][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 Microbiol21:811–821 [CrossRef][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 Bacteriol174:2493–2500[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 Chem279:12588–12597 [CrossRef][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. Methods25:402–408 [CrossRef][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 Bacteriol175:2221–2228[PubMed]
    [Google Scholar]
  25. Nordström K..( 2006;). Plasmid R1 – replication and its control. Plasmid55:1–26 [CrossRef][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 Res35:Web Server issueW433–W437 [CrossRef][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 Bacteriol184:3957–3964 [CrossRef][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 Bacteriol188:3582–3588 [CrossRef][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 Lett310:112–119 [CrossRef][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 Chem280:15084–15096 [CrossRef][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. Genetics184:947–958 [CrossRef][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 Biol218:119–128 [CrossRef][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 Bacteriol185:1616–1623 [CrossRef][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 Biol277:309–316 [CrossRef][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 Cells3:331–345 [CrossRef][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 Rev73:481–509 [CrossRef][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 Bacteriol191:1695–1702 [CrossRef][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 Bacteriol188:507–514 [CrossRef][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 Microbiol54:769–782 [CrossRef][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 Microbiol85:602–617 [CrossRef][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 Bacteriol188:6611–6621 [CrossRef][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 Bacteriol189:5885–5894 [CrossRef][PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.071738-0
Loading
/content/journal/micro/10.1099/mic.0.071738-0
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error