1887

Abstract

Bacterial gene regulation is controlled by complex regulatory cascades which integrate input environmental signals and adapt specific and adequate output cellular responses. These complex networks are far from being elucidated, in particular in . In the present study, we developed bacterial two-hybrid genome fragment libraries of the PAO1 strain to identify potential partners involved in the HptB/HsbR/HsbA pathway. This powerful tool, validated by the interaction previously described between HsbR and HsbA proteins, allowed us to demonstrate that the HsbR response regulator dimerizes through its PP2C/ATPase C-terminal effector domain, an observation further confirmed by pull-down experiments. This will also allow us to identify further new partners in this cascade.

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2012-08-01
2024-04-20
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References

  1. Bordi C., de Bentzmann S. ( 2011). Hacking into bacterial biofilms: a new therapeutic challenge. Ann Intensive Care 1:19 [View Article][PubMed]
    [Google Scholar]
  2. Bordi C., Lamy M. C., Ventre I., Termine E., Hachani A., Fillet S., Roche B., Bleves S., Méjean V. & other authors ( 2010). Regulatory RNAs and the HptB/RetS signalling pathways fine-tune Pseudomonas aeruginosa pathogenesis. Mol Microbiol 76:1427–1443 [View Article][PubMed]
    [Google Scholar]
  3. Brencic A., McFarland K. A., McManus H. R., Castang S., Mogno I., Dove S. L., Lory S. ( 2009). The GacS/GacA signal transduction system of Pseudomonas aeruginosa acts exclusively through its control over the transcription of the RsmY and RsmZ regulatory small RNAs. Mol Microbiol 73:434–445 [View Article][PubMed]
    [Google Scholar]
  4. Burrowes E., Baysse C., Adams C., O’Gara F. ( 2006). Influence of the regulatory protein RsmA on cellular functions in Pseudomonas aeruginosa PAO1, as revealed by transcriptome analysis. Microbiology 152:405–418 [View Article][PubMed]
    [Google Scholar]
  5. Clarke L., Carbon J. ( 1976). A colony bank containing synthetic Col El hybrid plasmids representative of the entire E. coli genome. Cell 9:91–99 [View Article][PubMed]
    [Google Scholar]
  6. De N., Navarro M. V., Raghavan R. V., Sondermann H. ( 2009). Determinants for the activation and autoinhibition of the diguanylate cyclase response regulator WspR. J Mol Biol 393:619–633 [View Article][PubMed]
    [Google Scholar]
  7. Delumeau O., Dutta S., Brigulla M., Kuhnke G., Hardwick S. W., Völker U., Yudkin M. D., Lewis R. J. ( 2004). Functional and structural characterization of RsbU, a stress signaling protein phosphatase 2C. J Biol Chem 279:40927–40937 [View Article][PubMed]
    [Google Scholar]
  8. Domain F., Bina X. R., Levy S. B. ( 2007). Transketolase A, an enzyme in central metabolism, derepresses the marRAB multiple antibiotic resistance operon of Escherichia coli by interaction with MarR. Mol Microbiol 66:383–394 [View Article][PubMed]
    [Google Scholar]
  9. Fields S., Song O. ( 1989). A novel genetic system to detect protein–protein interactions. Nature 340:245–246 [View Article][PubMed]
    [Google Scholar]
  10. Gao R., Stock A. M. ( 2010). Molecular strategies for phosphorylation-mediated regulation of response regulator activity. Curr Opin Microbiol 13:160–167 [View Article][PubMed]
    [Google Scholar]
  11. Goodman A. L., Kulasekara B., Rietsch A., Boyd D., Smith R. S., Lory S. ( 2004). A signaling network reciprocally regulates genes associated with acute infection and chronic persistence in Pseudomonas aeruginosa . Dev Cell 7:745–754 [View Article][PubMed]
    [Google Scholar]
  12. Handford J. I., Ize B., Buchanan G., Butland G. P., Greenblatt J., Emili A., Palmer T. ( 2009). Conserved network of proteins essential for bacterial viability. J Bacteriol 191:4732–4749 [View Article][PubMed]
    [Google Scholar]
  13. Heurlier K., Williams F., Heeb S., Dormond C., Pessi G., Singer D., Cámara M., Williams P., Haas D. ( 2004). Positive control of swarming, rhamnolipid synthesis, and lipase production by the posttranscriptional RsmA/RsmZ system in Pseudomonas aeruginosa PAO1. J Bacteriol 186:2936–2945 [View Article][PubMed]
    [Google Scholar]
  14. Hsu J. L., Chen H. C., Peng H. L., Chang H. Y. ( 2008). Characterization of the histidine-containing phosphotransfer protein B-mediated multistep phosphorelay system in Pseudomonas aeruginosa PAO1. J Biol Chem 283:9933–9944 [View Article][PubMed]
    [Google Scholar]
  15. Karimova G., Pidoux J., Ullmann A., Ladant D. ( 1998). A bacterial two-hybrid system based on a reconstituted signal transduction pathway. Proc Natl Acad Sci U S A 95:5752–5756 [View Article][PubMed]
    [Google Scholar]
  16. Karimova G., Ullmann A., Ladant D. ( 2001). Protein-protein interaction between Bacillus stearothermophilus tyrosyl-tRNA synthetase subdomains revealed by a bacterial two-hybrid system. J Mol Microbiol Biotechnol 3:73–82[PubMed]
    [Google Scholar]
  17. Karna S. L., Zogaj X., Barker J. R., Seshu J., Dove S. L., Klose K. E. ( 2010). A bacterial two-hybrid system that utilizes Gateway cloning for rapid screening of protein-protein interactions. Biotechniques 49:831–833 [View Article][PubMed]
    [Google Scholar]
  18. Kay E., Humair B., Dénervaud V., Riedel K., Spahr S., Eberl L., Valverde C., Haas D. ( 2006). Two GacA-dependent small RNAs modulate the quorum-sensing response in Pseudomonas aeruginosa . J Bacteriol 188:6026–6033 [View Article][PubMed]
    [Google Scholar]
  19. Kleinschnitz E. M., Heichlinger A., Schirner K., Winkler J., Latus A., Maldener I., Wohlleben W., Muth G. ( 2011). Proteins encoded by the mre gene cluster in Streptomyces coelicolor A3(2) cooperate in spore wall synthesis. Mol Microbiol 79:1367–1379 [View Article][PubMed]
    [Google Scholar]
  20. Klepp L. I., Soria M., Blanco F. C., Bianco M. V., Santangelo M. P., Cataldi A. A., Bigi F. ( 2009). Identification of two proteins that interact with the Erp virulence factor from Mycobacterium tuberculosis by using the bacterial two-hybrid system. BMC Mol Biol 10:3 [View Article][PubMed]
    [Google Scholar]
  21. Pessi G., Williams F., Hindle Z., Heurlier K., Holden M. T., Cámara M., Haas D., Williams P. ( 2001). The global posttranscriptional regulator RsmA modulates production of virulence determinants and N-acylhomoserine lactones in Pseudomonas aeruginosa . J Bacteriol 183:6676–6683 [View Article][PubMed]
    [Google Scholar]
  22. Rahme L. G., Ausubel F. M., Cao H., Drenkard E., Goumnerov B. C., Lau G. W., Mahajan-Miklos S., Plotnikova J., Tan M. W. & other authors ( 2000). Plants and animals share functionally common bacterial virulence factors. Proc Natl Acad Sci U S A 97:8815–8821 [View Article][PubMed]
    [Google Scholar]
  23. Reimmann C., Beyeler M., Latifi A., Winteler H., Foglino M., Lazdunski A., Haas D. ( 1997). The global activator GacA of Pseudomonas aeruginosa PAO positively controls the production of the autoinducer N-butyryl-homoserine lactone and the formation of the virulence factors pyocyanin, cyanide, and lipase. Mol Microbiol 24:309–319 [View Article][PubMed]
    [Google Scholar]
  24. Roucourt B., Lecoutere E., Chibeu A., Hertveldt K., Volckaert G., Lavigne R. ( 2009). A procedure for systematic identification of bacteriophage–host interactions of P. aeruginosa phages. Virology 387:50–58 [View Article][PubMed]
    [Google Scholar]
  25. Thibodeau S. A., Fang R., Joung J. K. ( 2004). High-throughput beta-galactosidase assay for bacterial cell-based reporter systems. Biotechniques 36:410–415[PubMed]
    [Google Scholar]
  26. Ventre I., Goodman A. L., Vallet-Gely I., Vasseur P., Soscia C., Molin S., Bleves S., Lazdunski A., Lory S., Filloux A. ( 2006). Multiple sensors control reciprocal expression of Pseudomonas aeruginosa regulatory RNA and virulence genes. Proc Natl Acad Sci U S A 103:171–176 [View Article][PubMed]
    [Google Scholar]
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