The Journal of Microbiology

, 47:572

Ligand-receptor recognition for activation of quorum sensing in Staphylococcus aureus

Article

Abstract

The accessory gene regulator (agr) locus controls many of the virulence toxins involved in Staphylococcus aureus pathogenesis, and can be divided into four specificity groups. AgrC is the only group-specific receptor to mediate both intra-group activation and inter-group inhibition. We studied the ligand-receptor recognition of the agr system in depth by using a luciferase reporter system to identify the key residues responsible for AgrC activation in two closely related agr groups, AgrC-I, and AgrC-IV. Fusion PCR and site-directed mutagenesis were used to screen for functional residues of AgrC. Our data suggest that for AgrC-IV activation, residue 101 is critical for activating the receptor. In contrast, the key residues for the activation of AgrC-I are located at residues 49∼59, 107, and 116. However, three residue changes, T101A, V107S, I116S, are sufficient to convert the AIP recognizing specificity from AgrC-IV to AgrC-I.

Keywords

histidine protein kinases ligands protein structure quorum sensing transmembrane receptors two-component system 

References

  1. Ballesteros, J.A., L. Shi, and J.A. Javitch. 2001. Structural mimicry in G protein-coupled receptors: implications of the high-resolution structure of rhodopsin for structure-function analysis of rhodopsin-like receptors. Mol. Pharmacol. 60, 1–19.PubMedGoogle Scholar
  2. Betts, M.J. and R.B. Russell. 2003. Amino acid properties and consequences of substitutions, p. 289–316. In I.C. Gray and M.R. Barnes (eds.), Bioinformatics for geneticists.Google Scholar
  3. Chen, F.J., A. Cockayne, K. Winzer, W.C. Chan, H.J. Lo, and P. Williams. 2004. Probing AgrC-AIP interactions in Staphylococcus aureus. Cell-Cell Communication in Bacteria (2nd). Abstr. 76A. American Society for Microbiology, Banff, Alberta, Canada.Google Scholar
  4. Geisinger, E., E.A. George, T.W. Muir, and R.P. Novick. 2008. Identification of ligand specificity determinants in AgrC, the Staphylococcus aureus quorum-sensing receptor. J. Biol. Chem. 283, 8930–8938.CrossRefPubMedGoogle Scholar
  5. Geisinger, E., T.W. Muir, and R.P. Novick. 2009. agr Receptor mutants reveal distinct modes of inhibition by staphylococcal autoinducing peptides. Proc. Natl. Acad. Sci. USA 106, 1216–1221.CrossRefPubMedGoogle Scholar
  6. Gether, U. 2000. Uncovering molecular mechanisms involved in activation of G protein-coupled receptors. Endocr. Rev. 21, 90–113.CrossRefPubMedGoogle Scholar
  7. Gether, U. and B.K. Kobilka. 1998. G protein-coupled receptors. II. mechanism of agonist activation. J. Biol. Chem. 273, 17979–17982.CrossRefPubMedGoogle Scholar
  8. Greenspan, N.S. and E. Di Cera. 1999. Defining epitopes: It’s not as easy as it seems. Nat. Biotechnol. 17, 936–937.CrossRefPubMedGoogle Scholar
  9. Grkovic, S., M.H. Brown, K.M. Hardie, N. Firth, and R.A. Skurray. 2003. Stable low-copy-number Staphylococcus aureus shuttle vectors. Microbiology 149, 785–794.CrossRefPubMedGoogle Scholar
  10. Janzon, L. and S. Arvidson. 1990. The role of the delta-lysin gene (hld) in the regulation of virulence genes by the accessory gene regulator (agr) in Staphylococcus aureus. EMBO J. 9, 1391–1399.PubMedGoogle Scholar
  11. Janzon, L., S. Lofdahl, and S. Arvidson. 1989. Identification and nucleotide sequence of the delta-lysin gene, hld, adjacent to the accessory gene regulator (agr) of Staphylococcus aureus. Mol. Gen. Genet. 219, 480–485.CrossRefPubMedGoogle Scholar
  12. Jarraud, S., G.J. Lyon, A.M. Figueiredo, L. Gerard, F. Vandenesch, J. Etienne, T.W. Muir, and R.P. Novick. 2000. Exfoliatin-producing strains define a fourth agr specificity group in Staphylococcus aureus. J. Bacteriol. 182, 6517–6522.CrossRefPubMedGoogle Scholar
  13. Jensen, R.O., K. Winzer, S.R. Clarke, W.C. Chan, and P. Williams. 2008. Differential recognition of Staphylococcus aureus quorum-sensing signals depends on both extracellular loops 1 and 2 of the transmembrane sensor AgrC. J. Mol. Biol. 381, 300–309.CrossRefPubMedGoogle Scholar
  14. Ji, G., R.C. Beavis, and R.P. Novick. 1995. Cell density control of staphylococcal virulence mediated by an octapeptide pheromone. Proc. Natl. Acad. Sci. USA 92, 12055–12059.CrossRefPubMedGoogle Scholar
  15. Ji, G., R. Beavis, and R.P. Novick. 1997. Bacterial interference caused by autoinducing peptide variants. Science 276, 2027–2030.CrossRefPubMedGoogle Scholar
  16. Koenig, R.L., J.L. Ray, S.J. Maleki, M.S. Smeltzer, and B.K. Hurlburt. 2004. Staphylococcus aureus AgrA binding to the RNAIII-agr regulatory region. J. Bacteriol. 186, 7549–7555.CrossRefPubMedGoogle Scholar
  17. Koretke, K.K., A.N. Lupas, P.V. Warren, M. Rosenberg, and J.R. Brown. 2000. Evolution of two-component signal transduction. Mol. Biol. Evol. 17, 1956–1970.PubMedGoogle Scholar
  18. Krogh, A., B. Larsson, G. von Heijne, and E.L. Sonnhammer. 2001. Predicting transmembrane protein topology with a hidden Markov model: application to complete genomes. J. Mol. Biol. 305, 567–580.CrossRefPubMedGoogle Scholar
  19. Lawson, Z. and M. Wheatley. 2004. The third extracellular loop of G protein-coupled receptors: more than just a linker between two important transmembrane helices. Biochem. Soc. Trans. 32, 1048–1050.CrossRefPubMedGoogle Scholar
  20. Lina, G., S. Jarraud, G. Ji, T. Greenland, A. Pedraza, J. Etienne, R.P. Novick, and F. Vandenesch. 1998. Transmembrane topology and histidine protein kinase activity of AgrC, the agr signal receptor in Staphylococcus aureus. Mol. Microbiol. 28, 655–662.CrossRefPubMedGoogle Scholar
  21. Lyon, G.J., P. Mayville, T.W. Muir, and R.P. Novick. 2000. Rational design of a global inhibitor of the virulence response in Staphylococcus aureus, based in part on localization of the site of inhibition to the receptor-histidine kinase, AgrC. Proc. Natl. Acad. Sci. USA 97, 13330–13335.CrossRefPubMedGoogle Scholar
  22. Lyon, G.J. and R.P. Novick. 2004. Peptide signaling in Staphylococcus aureus and other Gram-positive bacteria. Peptides 25, 1389–1403.CrossRefPubMedGoogle Scholar
  23. Lyon, G.J., J.S. Wright, T.W. Muir, and R.P. Novick. 2002. Key determinants of receptor activation in the agr autoinducing peptides of Staphylococcus aureus. Biochemistry 41, 10095–10104.CrossRefPubMedGoogle Scholar
  24. McDowell, P., Z. Affas, C. Reynolds, M.T. Holden, S.J. Wood, S. Saint, A. Cockayne, P.J. Hill, C.E. Dodd, B.W. Bycroft, W.C. Chan, and P. Williams. 2001. Structure, activity and evolution of the group I thiolactone peptide quorum-sensing system of Staphylococcus aureus. Mol. Microbiol. 41, 503–512.CrossRefGoogle Scholar
  25. Morfeldt, E., L. Janzon, S. Arvidson, and S. Lofdahl. 1988. Cloning of a chromosomal locus (exp) which regulates the expression of several exoprotein genes in Staphylococcus aureus. Mol. Gen. Genet. 211, 435–440.CrossRefPubMedGoogle Scholar
  26. Norrander, J., T. Kempe, and J. Messing. 1983. Construction of improved M13 vectors using oligodeoxynucleotide-directed mutagenesis. Gene 26, 101–106.CrossRefPubMedGoogle Scholar
  27. Novick, R.P. 1991. Genetic systems in staphylococci. Methods Enzymol. 204, 587–636.CrossRefPubMedGoogle Scholar
  28. Novick, R.P., S.J. Projan, J. Kornblum, H.F. Ross, G. Ji, B. Kreiswirth, F. Vandenesch, and S. Moghazeh. 1995. The agr P2 operon: an autocatalytic sensory transduction system in Staphylococcus aureus. Mol. Gen. Genet. 248, 446–458.CrossRefPubMedGoogle Scholar
  29. Novick, R.P., H.F. Ross, S.J. Projan, J. Kornblum, B. Kreiswirth, and S. Moghazeh. 1993. Synthesis of staphylococcal virulence factors is controlled by a regulatory RNA molecule. EMBO J. 12, 3967–3975.PubMedGoogle Scholar
  30. Otto, M. 2001. Staphylococcus aureus and Staphylococcus epidermidis peptide pheromones produced by the accessory gene regulator agr system. Peptides 22, 1603–1608.CrossRefPubMedGoogle Scholar
  31. Peng, H.L., R.P. Novick, B. Kreiswirth, J. Kornblum, and P. Schlievert. 1988. Cloning, characterization, and sequencing of an accessory gene regulator (agr) in Staphylococcus aureus. J. Bacteriol. 170, 4365–4372.PubMedGoogle Scholar
  32. Qazi, S.N., E. Counil, J. Morrissey, C.E. Rees, A. Cockayne, K. Winzer, W.C. Chan, P. Williams, and P.J. Hill. 2001. agr Expression precedes escape of internalized Staphylococcus aureus from the host endosome. Infect. Immun. 69, 7074–7082.CrossRefPubMedGoogle Scholar
  33. Recsei, P., B. Kreiswirth, M. O’Reilly, P. Schlievert, A. Gruss, and R.P. Novick. 1986. Regulation of exoprotein gene expression in Staphylococcus aureus by agr. Mol. Gen. Genet. 202, 58–61.CrossRefPubMedGoogle Scholar
  34. Schenk, S. and R.A. Laddaga. 1992. Improved method for electroporation of Staphylococcus aureus. FEMS Microbiol. Lett. 73, 133–138.CrossRefPubMedGoogle Scholar
  35. van Leeuwen, W., W. van Nieuwenhuizen, C. Gijzen, H. Verbrugh, and A. van Belkum. 2000. Population studies of methicillin-resistant and -sensitive Staphylococcus aureus strains reveal a lack of variability in the agrD gene, encoding a staphylococcal autoinducer peptide. J. Bacteriol. 182, 5721–5729.CrossRefPubMedGoogle Scholar
  36. Wolanin, P.M., P.A. Thomason, and J.B. Stock. 2002. Histidine protein kinases: key signal transducers outside the animal kingdom. Genome Biol. 3, reviews3013.Google Scholar
  37. Wright, J.S., 3rd, G.J. Lyon, E.A. George, T.W. Muir, and R.P. Novick. 2004. Hydrophobic interactions drive ligand-receptor recognition for activation and inhibition of staphylococcal quorum sensing. Proc. Natl. Acad. Sci. USA 101, 16168–16173.CrossRefPubMedGoogle Scholar

Copyright information

© The Microbiological Society of Korea and Springer Berlin Heidelberg 2009

Authors and Affiliations

  1. 1.Division of Infectious DiseasesNational Health Research InstitutesZhunan Town, Miaoli CountyTaiwan

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