Two Group A Streptococcal Peptide Pheromones Act through Opposing Rgg Regulators to Control Biofilm Development

English version České info

Streptococcus pyogenes (Group A Streptococcus, GAS) is an important human commensal that occasionally causes localized infections and less frequently causes severe invasive disease with high mortality rates. How GAS regulates expression of factors used to colonize the host and avoid immune responses remains poorly understood. Intercellular communication is an important means by which bacteria coordinate gene expression to defend against host assaults and competing bacteria, yet no conserved cell-to-cell signaling system has been elucidated in GAS. Encoded within the GAS genome are four rgg-like genes, two of which (rgg2 and rgg3) have no previously described function. We tested the hypothesis that rgg2 or rgg3 rely on extracellular peptides to control target-gene regulation. We found that Rgg2 and Rgg3 together tightly regulate two linked genes encoding new peptide pheromones. Rgg2 activates transcription of and is required for full induction of the pheromone genes, while Rgg3 plays an antagonistic role and represses pheromone expression. The active pheromone signals, termed SHP2 and SHP3, are short and hydrophobic (DI[I/L]IIVGG), and, though highly similar in sequence, their ability to disrupt Rgg3-DNA complexes were observed to be different, indicating that specificity and differential activation of promoters are characteristics of the Rgg2/3 regulatory circuit. SHP-pheromone signaling requires an intact oligopeptide permease (opp) and a metalloprotease (eep), supporting the model that pro-peptides are secreted, processed to the mature form, and subsequently imported to the cytoplasm to interact directly with the Rgg receptors. At least one consequence of pheromone stimulation of the Rgg2/3 pathway is increased biogenesis of biofilms, which counteracts negative regulation of biofilms by RopB (Rgg1). These data provide the first demonstration that Rgg-dependent quorum sensing functions in GAS and substantiate the role that Rggs play as peptide receptors across the Firmicute phylum.

Vyšlo v časopise: Two Group A Streptococcal Peptide Pheromones Act through Opposing Rgg Regulators to Control Biofilm Development. PLoS Pathog 7(8): e32767. doi:10.1371/journal.ppat.1002190
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1002190

Souhrn

Zdroje

1. RobinsonCJBohannanBJYoungVB 2010 From structure to function: the ecology of host-associated microbial communities. Microbiol Mol Biol Rev 74 453 476

2. WilsonM 2005 Microbial inhabitants of humans : their ecology and role in health and disease. New York Cambridge University Press. xviii, 455 p

3. HughesDTSperandioV 2008 Inter-kingdom signalling: communication between bacteria and their hosts. Nat Rev Microbiol 6 111 120

4. NgWLBasslerBL 2009 Bacterial quorum-sensing network architectures. Annu Rev Genet 43 197 222

5. FederleMJBasslerBL 2003 Interspecies communication in bacteria. J Clin Invest 112 1291 1299

6. ClaverysJPHavarsteinLS 2007 Cannibalism and fratricide: mechanisms and raisons d'etre. Nat Rev Microbiol 5 219 229

7. StraightPDKolterR 2009 Interspecies chemical communication in bacterial development. Annu Rev Microbiol 63 99 118

8. IrieYParsekMR 2008 Quorum sensing and microbial biofilms. Curr Top Microbiol Immunol 322 67 84

9. PechèreJCKaplanEL 2004 Streptococcal pharyngitis : optimal management. Basel ; New York Karger. xii, 220 p

10. MartinJMGreenMBarbadoraKAWaldER 2004 Group A streptococci among school-aged children: clinical characteristics and the carrier state. Pediatrics 114 1212 1219

11. CunninghamMW 2000 Pathogenesis of group A streptococcal infections. Clin Microbiol Rev 13 470 511

12. StevensDLKaplanEL 2000 Streptococcal infections : clinical aspects, microbiology, and molecular pathogenesis. New York Oxford University Press. xv, 449 p

13. KaplanEL 1980 The group A streptococcal upper respiratory tract carrier state: an enigma. J Pediatr 97 337 345

14. DaltonTLScottJR 2004 CovS inactivates CovR and is required for growth under conditions of general stress in Streptococcus pyogenes. J Bacteriol 186 3928 3937

15. GryllosITran-WinklerHJChengMFChungHBolcomeR3rd 2008 Induction of group A Streptococcus virulence by a human antimicrobial peptide. Proc Natl Acad Sci U S A 105 16755 16760

16. KietzmanCCCaparonMG 2010 CcpA and LacD.1 affect temporal regulation of Streptococcus pyogenes virulence genes. Infect Immun 78 241 252

17. MalkeHSteinerKMcShanWMFerrettiJJ 2006 Linking the nutritional status of Streptococcus pyogenes to alteration of transcriptional gene expression: the action of CodY and RelA. Int J Med Microbiol 296 259 275

18. KreikemeyerBMcIverKSPodbielskiA 2003 Virulence factor regulation and regulatory networks in Streptococcus pyogenes and their impact on pathogen-host interactions. Trends Microbiol 11 224 232

19. FontaineLBoutryCHenry de FrahanMDelplaceBFremauxC 2009 A novel pheromone quorum-sensing system controls the development of natural competence in Streptococcus thermophilus and Streptococcus salivarius. J Bacteriol 192 1444 54

20. IbrahimMGuillotAWessnerFAlgaronFBessetC 2007 Control of the transcription of a short gene encoding a cyclic peptide in Streptococcus thermophilus: a new quorum-sensing system? J Bacteriol 189 8844 8854

21. Mashburn-WarrenLMorrisonDAFederleMJ 2010 A novel double-tryptophan peptide pheromone controls competence in Streptococcus spp. via an Rgg regulator. Mol Microbiol 78 589 606

22. StephensonSMuellerCJiangMPeregoM 2003 Molecular analysis of Phr peptide processing in Bacillus subtilis. J Bacteriol 185 4861 4871

23. ZhangLGrayLNovickRPJiG 2002 Transmembrane topology of AgrB, the protein involved in the post-translational modification of AgrD in Staphylococcus aureus. J Biol Chem 277 34736 34742

24. HuiFMMorrisonDA 1991 Genetic transformation in Streptococcus pneumoniae: nucleotide sequence analysis shows comA, a gene required for competence induction, to be a member of the bacterial ATP-dependent transport protein family. J Bacteriol 173 372 381

25. AnFYSulavikMCClewellDB 1999 Identification and characterization of a determinant (eep) on the Enterococcus faecalis chromosome that is involved in production of the peptide sex pheromone cAD1. J Bacteriol 181 5915 5921

26. Lanigan-GerdesSDooleyANFaullKFLazazzeraBA 2007 Identification of subtilisin, Epr and Vpr as enzymes that produce CSF, an extracellular signalling peptide of Bacillus subtilis. Mol Microbiol 65 1321 1333

27. OttoMSussmuthRJungGGotzF 1998 Structure of the pheromone peptide of the Staphylococcus epidermidis agr system. FEBS Lett 424 89 94

28. MayvillePJiGBeavisRYangHGogerM 1999 Structure-activity analysis of synthetic autoinducing thiolactone peptides from Staphylococcus aureus responsible for virulence. Proc Natl Acad Sci U S A 96 1218 1223

29. ThoendelMHorswillAR 2009 Identification of Staphylococcus aureus AgrD residues required for autoinducing peptide biosynthesis. J Biol Chem 284 21828 21838

30. MagnusonRSolomonJGrossmanAD 1994 Biochemical and genetic characterization of a competence pheromone from B. subtilis. Cell 77 207 216

31. AnsaldiMMaroltDStebeTMandic-MulecIDubnauD 2002 Specific activation of the Bacillus quorum-sensing systems by isoprenylated pheromone variants. Mol Microbiol 44 1561 1573

32. NakayamaJCaoYHoriiTSakudaSAkkermansAD 2001 Gelatinase biosynthesis-activating pheromone: a peptide lactone that mediates a quorum sensing in Enterococcus faecalis. Mol Microbiol 41 145 154

33. DetmersFJLanfermeijerFCAbeleRJackRWTampeR 2000 Combinatorial peptide libraries reveal the ligand-binding mechanism of the oligopeptide receptor OppA of Lactococcus lactis. Proc Natl Acad Sci U S A 97 12487 12492

34. DoevenMKAbeleRTampeRPoolmanB 2004 The binding specificity of OppA determines the selectivity of the oligopeptide ATP-binding cassette transporter. J Biol Chem 279 32301 32307

35. SolomonJSuLShynSGrossmanAD 2003 Isolation and characterization of mutants of the Bacillus subtilis oligopeptide permease with altered specificity of oligopeptide transport. J Bacteriol 185 6425 6433

36. DeclerckNBouillautLChaixDRuganiNSlamtiL 2007 Structure of PlcR: Insights into virulence regulation and evolution of quorum sensing in Gram-positive bacteria. Proc Natl Acad Sci U S A 104 18490 18495

37. Rocha-EstradaJAceves-DiezAGuarnerosGde la TorreM 2010 The RNPP family of quorum-sensing proteins in Gram-positive bacteria. Appl Microbiol Biotechnol 87 913 923

38. SulavikMCTardifGClewellDB 1992 Identification of a gene, rgg, which regulates expression of glucosyltransferase and influences the Spp phenotype of Streptococcus gordonii Challis. J Bacteriol 174 3577 3586

39. KelleyLASternbergMJ 2009 Protein structure prediction on the Web: a case study using the Phyre server. Nat Protoc 4 363 371

40. ZhangY 2008 I-TASSER server for protein 3D structure prediction. BMC Bioinformatics 9 40

41. BelotserkovskyIBaruchMPeerADovERavinsM 2009 Functional analysis of the quorum-sensing streptococcal invasion locus (sil). PLoS Pathog 5 e1000651

42. Hidalgo-GrassCRavinsMDan-GoorMJaffeJMosesAE 2002 A locus of group A Streptococcus involved in invasive disease and DNA transfer. Mol Microbiol 46 87 99

43. ChausseeMSSylvaGLSturdevantDESmootLMGrahamMR 2002 Rgg influences the expression of multiple regulatory loci to coregulate virulence factor expression in Streptococcus pyogenes. Infect Immun 70 762 770

44. HollandsAAzizRKKansalRKotbMNizetV 2008 A naturally occurring mutation in ropB suppresses SpeB expression and reduces M1T1 group A streptococcal systemic virulence. PLoS One 3 e4102

45. LyonWRGibsonCMCaparonMG 1998 A role for trigger factor and an rgg-like regulator in the transcription, secretion and processing of the cysteine proteinase of Streptococcus pyogenes. Embo J 17 6263 6275

46. NeelyMNLyonWRRunftDLCaparonM 2003 Role of RopB in growth phase expression of the SpeB cysteine protease of Streptococcus pyogenes. J Bacteriol 185 5166 5174

47. IbrahimMNicolasPBessieresPBolotinAMonnetV 2007 A genome-wide survey of short coding sequences in streptococci. Microbiology 153 3631 3644

48. McShanWMFerrettiJJKarasawaTSuvorovANLinS 2008 Genome sequence of a nephritogenic and highly transformable M49 strain of Streptococcus pyogenes. J Bacteriol 190 7773 7785

49. ToukokiCGoldKMMcIverKSEichenbaumZ 2010 MtsR is a dual regulator that controls virulence genes and metabolic functions in addition to metal homeostasis in the group A streptococcus. Mol Microbiol 76 971 989

50. McShanWMMcLaughlinRENordstrandAFerrettiJJ 2004 Vectors containing streptococcal bacteriophage integrases for site-specific gene insertion Methods in Cell Science 20 51 57

51. van de RijnIKesslerRE 1980 Growth characteristics of group A streptococci in a new chemically defined medium. Infect Immun 27 444 448

52. FleuchotBGittonCGuillotAVidicJNicolasP 2011 Rgg proteins associated with internalized small hydrophobic peptides: a new quorum-sensing mechanism in streptococci. Mol Microbiol 80 1102 19

53. SamenUMEikmannsBJReinscheidDJ 2006 The transcriptional regulator RovS controls the attachment of Streptococcus agalactiae to human epithelial cells and the expression of virulence genes. Infect Immun 74 5625 5635

54. VickermanMMWangMBakerLJ 2003 An amino acid change near the carboxyl terminus of the Streptococcus gordonii regulatory protein Rgg affects its abilities to bind DNA and influence expression of the glucosyltransferase gene gtfG. Microbiology 149 399 406

55. HilesIDHigginsCF 1986 Peptide uptake by Salmonella typhimurium. The periplasmic oligopeptide-binding protein. Eur J Biochem 158 561 567

56. GoodellEWHigginsCF 1987 Uptake of cell wall peptides by Salmonella typhimurium and Escherichia coli. J Bacteriol 169 3861 3865

57. LazazzeraBASolomonJMGrossmanAD 1997 An exported peptide functions intracellularly to contribute to cell density signaling in B. subtilis. Cell 89 917 925

58. SlamtiLLereclusD 2002 A cell-cell signaling peptide activates the PlcR virulence regulon in bacteria of the Bacillus cereus group. Embo J 21 4550 4559

59. LeonardBAPodbielskiAHedbergPJDunnyGM 1996 Enterococcus faecalis pheromone binding protein, PrgZ, recruits a chromosomal oligopeptide permease system to import sex pheromone cCF10 for induction of conjugation. Proc Natl Acad Sci U S A 93 260 264

60. PodbielskiALeonardBA 1998 The group A streptococcal dipeptide permease (Dpp) is involved in the uptake of essential amino acids and affects the expression of cysteine protease. Mol Microbiol 28 1323 1334

61. PodbielskiAPohlBWoischnikMKornerCSchmidtKH 1996 Molecular characterization of group A streptococcal (GAS) oligopeptide permease (opp) and its effect on cysteine protease production. Mol Microbiol 21 1087 1099

62. AnFYClewellDB 2002 Identification of the cAD1 sex pheromone precursor in Enterococcus faecalis. J Bacteriol 184 1880 1887

63. ChandlerJRDunnyGM 2008 Characterization of the sequence specificity determinants required for processing and control of sex pheromone by the intramembrane protease Eep and the plasmid-encoded protein PrgY. J Bacteriol 190 1172 1183

64. VickermanMMFlannaganSEJesionowskiAMBrossardKAClewellDB 2010 A genetic determinant in Streptococcus gordonii Challis encodes a peptide with activity similar to that of enterococcal sex pheromone cAM373, which facilitates intergeneric DNA transfer. J Bacteriol 192 2535 2545

65. DenhamELWardPNLeighJA 2008 Lipoprotein signal peptides are processed by Lsp and Eep of Streptococcus uberis. J Bacteriol 190 4641 4647

66. CserzoMWallinESimonIvon HeijneGElofssonA 1997 Prediction of transmembrane alpha-helices in prokaryotic membrane proteins: the dense alignment surface method. Protein Eng 10 673 676

67. KaratanEWatnickP 2009 Signals, regulatory networks, and materials that build and break bacterial biofilms. Microbiol Mol Biol Rev 73 310 347

68. LembkeCPodbielskiAHidalgo-GrassCJonasLHanskiE 2006 Characterization of biofilm formation by clinically relevant serotypes of group A streptococci. Appl Environ Microbiol 72 2864 2875

69. CourtneyHSOfekIPenfoundTNizetVPenceMA 2009 Relationship between expression of the family of M proteins and lipoteichoic acid to hydrophobicity and biofilm formation in Streptococcus pyogenes. PLoS One 4 e4166

70. ThenmozhiRBalajiKKumarRRaoTSPandianSK 2011 Characterization of biofilms in different clinical M serotypes of Streptococcus pyogenes. J Basic Microbiol 51 196 204

71. DoernCDRobertsALHongWNelsonJLukomskiS 2009 Biofilm formation by group A Streptococcus: a role for the streptococcal regulator of virulence (Srv) and streptococcal cysteine protease (SpeB). Microbiology 155 46 52

72. ThoendelMHorswillAR 2010 Biosynthesis of peptide signals in gram-positive bacteria. Adv Appl Microbiol 71 91 112

73. NovickRPGeisingerE 2008 Quorum sensing in staphylococci. Annu Rev Genet 42 541 564

74. GoharMFaegriKPerchatSRavnumSOkstadOA 2008 The PlcR virulence regulon of Bacillus cereus. PLoS One 3 e2793

75. PottathilMLazazzeraBA 2003 The extracellular Phr peptide-Rap phosphatase signaling circuit of Bacillus subtilis. Front Biosci 8 d32 45

76. ClaverysJPPrudhommeMMartinB 2006 Induction of competence regulons as a general response to stress in gram-positive bacteria. Annu Rev Microbiol 60 451 475

77. DunnyGM 2007 The peptide pheromone-inducible conjugation system of Enterococcus faecalis plasmid pCF10: cell-cell signalling, gene transfer, complexity and evolution. Philos Trans R Soc Lond B Biol Sci 362 1185 1193

78. EranYGetterYBaruchMBelotserkovskyIPadalonG 2007 Transcriptional regulation of the sil locus by the SilCR signalling peptide and its implications on group A streptococcus virulence. Mol Microbiol 63 1209 1222

79. ConleyJOlsonMECookLSCeriHPhanV 2003 Biofilm formation by group a streptococci: is there a relationship with treatment failure? J Clin Microbiol 41 4043 4048

80. BekalSGaudreauCLaurenceRASimoneauERaynalL 2006 Streptococcus pseudoporcinus sp. nov., a novel species isolated from the genitourinary tract of women. J Clin Microbiol 44 2584 2586

81. AltschulSFGishWMillerWMyersEWLipmanDJ 1990 Basic local alignment search tool. J Mol Biol 215 403 410

82. BiswasIScottJR 2003 Identification of rocA, a positive regulator of covR expression in the group A streptococcus. J Bacteriol 185 3081 3090

83. KizyAENeelyMN 2009 First Streptococcus pyogenes signature-tagged mutagenesis screen identifies novel virulence determinants. Infect Immun 77 1854 1865

84. MillerJDNeelyMN 2005 Large-scale screen highlights the importance of capsule for virulence in the zoonotic pathogen Streptococcus iniae. Infect Immun 73 921 934

85. ShiKBrownCKGuZYKozlowiczBKDunnyGM 2005 Structure of peptide sex pheromone receptor PrgX and PrgX/pheromone complexes and regulation of conjugation in Enterococcus faecalis. Proc Natl Acad Sci U S A 102 18596 18601

86. Howell-AdamsBSeifertHS 2000 Molecular models accounting for the gene conversion reactions mediating gonococcal pilin antigenic variation. Mol Microbiol 37 1146 1158

87. MirouxBWalkerJE 1996 Over-production of proteins in Escherichia coli: mutant hosts that allow synthesis of some membrane proteins and globular proteins at high levels. J Mol Biol 260 289 298

88. ClaverysJPDintilhacAPestovaEVMartinBMorrisonDA 1995 Construction and evaluation of new drug-resistance cassettes for gene disruption mutagenesis in Streptococcus pneumoniae, using an ami test platform. Gene 164 123 128

89. DegnanBAFontaineMCDoebereinerAHLeeJJMastroeniP 2000 Characterization of an isogenic mutant of Streptococcus pyogenes Manfredo lacking the ability to make streptococcal acid glycoprotein. Infect Immun 68 2441 2448

90. CharpentierEAntonAIBarryPAlfonsoBFangY 2004 Novel cassette-based shuttle vector system for gram-positive bacteria. Appl Environ Microbiol 70 6076 6085

91. HusmannLKScottJRLindahlGStenbergL 1995 Expression of the Arp protein, a member of the M protein family, is not sufficient to inhibit phagocytosis of Streptococcus pyogenes. Infect Immun 63 345 348

92. FoleyDASwartzentruberKGLavieAColleyKJ 2010 Structure and mutagenesis of neural cell adhesion molecule domains: evidence for flexibility in the placement of polysialic acid attachment sites. J Biol Chem 285 27360 27371

93. ManettiAGZingarettiCFalugiFCapoSBombaciM 2007 Streptococcus pyogenes pili promote pharyngeal cell adhesion and biofilm formation. Mol Microbiol 64 968 983

94. SimonDFerrettiJJ 1991 Electrotransformation of Streptococcus pyogenes with plasmid and linear DNA. FEMS Microbiol Lett 66 219 224

95. MaguinEDuwatPHegeTEhrlichDGrussA 1992 New thermosensitive plasmid for gram-positive bacteria. J Bacteriol 174 5633 5638