Inhibition of Competence Development, Horizontal Gene Transfer and Virulence in by a Modified Competence Stimulating Peptide

English version České info

Competence stimulating peptide (CSP) is a 17-amino acid peptide pheromone secreted by Streptococcus pneumoniae. Upon binding of CSP to its membrane-associated receptor kinase ComD, a cascade of signaling events is initiated, leading to activation of the competence regulon by the response regulator ComE. Genes encoding proteins that are involved in DNA uptake and transformation, as well as virulence, are upregulated. Previous studies have shown that disruption of key components in the competence regulon inhibits DNA transformation and attenuates virulence. Thus, synthetic analogues that competitively inhibit CSPs may serve as attractive drugs to control pneumococcal infection and to reduce horizontal gene transfer during infection. We performed amino acid substitutions on conserved amino acid residues of CSP1 in an effort to disable DNA transformation and to attenuate the virulence of S. pneumoniae. One of the mutated peptides, CSP1-E1A, inhibited development of competence in DNA transformation by outcompeting CSP1 in time and concentration-dependent manners. CSP1-E1A reduced the expression of pneumococcal virulence factors choline binding protein D (CbpD) and autolysin A (LytA) in vitro, and significantly reduced mouse mortality after lung infection. Furthermore, CSP1-E1A attenuated the acquisition of an antibiotic resistance gene and a capsule gene in vivo. Finally, we demonstrated that the strategy of using a peptide inhibitor is applicable to other CSP subtype, including CSP2. CSP1-E1A and CSP2-E1A were able to cross inhibit the induction of competence and DNA transformation in pneumococcal strains with incompatible ComD subtypes. These results demonstrate the applicability of generating competitive analogues of CSPs as drugs to control horizontal transfer of antibiotic resistance and virulence genes, and to attenuate virulence during infection by S. pneumoniae.

Vyšlo v časopise: Inhibition of Competence Development, Horizontal Gene Transfer and Virulence in by a Modified Competence Stimulating Peptide. PLoS Pathog 7(9): e32767. doi:10.1371/journal.ppat.1002241
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1002241

Souhrn

Zdroje

1. WeiserJN 2010 The pneumococcus: why a commensal misbehaves. J Mol Med 88 97 102

2. MitchellAMMitchellTJ 2010 Streptococcus pneumoniae: virulence factors and variation. Clin Microbiol Infect 16 411 418

3. KadiogluAWeiserJNPatonJCAndrewPW 2008 The role of Streptococcus pneumoniae virulence factors in host respiratory colonization and disease. Nat Rev Microbiol 6 288 301

4. CokerTRChanLSNewberrySJLimbosMASuttorpMJ 2010 Diagnosis, Microbial Epidemiology, and Antibiotic Treatment of Acute Otitis Media in Children: A Systematic Review. JAMA 304 2161 2169

5. LynchJP3rdZhanelGG 2010 Streptococcus pneumoniae: epidemiology and risk factors, evolution of antimicrobial resistance, and impact of vaccines. Curr Opin Pulm Med 16 217 225

6. LiñaresJArdanuyCPallaresRFenollA 2010 Changes in antimicrobial resistance, serotypes and genotypes in Streptococcus pneumoniae over a 30-year period. Clin Microbiol Infect 16 402 410

7. JohnsborgOHåvarsteinLS 2009 Regulation of natural genetic transformation and acquisition of transforming DNA in Streptococcus pneumoniae. FEMS Microbiol Rev 33 627 642

8. ClaverysJPMartinBPolardP 2009 The genetic transformation machinery: composition, localization, and mechanism. FEMS Microbiol Rev 33 643 656

9. MorrisonDALeeMS 2000 Regulation of competence for genetic transformation in Streptococcus pneumoniae: a link between quorum sensing and DNA processing genes. Res Microbiol 151 445 451

10. HavarsteinLSCoomaraswamyGMorrisonDA 1995 An unmodified heptadecapeptide pheromone induces competence for genetic transformation in Streptococcus pneumoniae. Proc Natl Acad Sci USA 92 11140 11144

11. PestovaEVHåvarsteinLSMorrisonDA 1996 Regulation of competence for genetic transformation in Streptococcus pneumoniae by an auto-induced peptide pheromone and a two-component regulatory system. Mol Microbiol 21 853 862

12. HåvarsteinLSGaustadPNesIFMorrisonDA 1996 Identification of the streptococcal competence-pheromone receptor. Mol Microbiol 21 863 869

13. MartinBGranadelCCampoNHénardVPrudhommeMClaverysJP 2010 Expression and maintenance of ComD-ComE, the two-component signal-transduction system that controls competence of Streptococcus pneumoniae. Mol Microbiol 75 1513 1528

14. PetersonSSungCKClineRDesaiBVSnesrudE 2004 Identification of competence pheromone responsive genes in Streptococcus pneumoniae. Mol Microbiol 51 1051 1070

15. LuoPLiHMorrisonDA 2003 ComX is a unique link between multiple quorum sensing outputs and competence in Streptococcus pneumoniae. Mol Microbiol 50 623 633

16. PozziGMasalaLLannelliFManganelliRHåvarsteinLS 1996 Competence for genetic transformation in encapsulated strains of Streptococcus pneumoniae: two allelic variants of the peptide pheromone. J Bacteriol 178 6087 6090

17. WhatmoreAMBarcusVADowsonCG 1999 Genetic diversity of the streptococcal competence (com) gene locus. J Bacteriol 181 3144 3154

18. IannelliFOggioniMRPozziG 2005 Sensor domain of histidine kinase ComD confers competence pherotype specificity in Streptoccoccus pneumoniae. FEMS Microbiol Lett 252 321 326

19. JohnsborgOKristiansenPEBlomqvistTHåvarsteinLS 2006 A Hydrophobic Patch in the Comptence-Stimulating Peptide, a Pneumococcal Competence Pheromone, Is Essential for Specificity and Biological Activity. J. Bacteriol 188 1744 1749

20. LauGWHaatajaSLonettoMKensitSEMarraA 2001 A functional genomic analysis of type 3 Streptococcus penumoniae virulence. Mol Microbiol 40 555 571

21. KowalkoJESebertME 2008 The Streptococcus pneumoniae competence regulatory system influences respiratory tract colonization. Infect Immun 76 3131 3140

22. IbrahimYMKerrARMcCluskeyJMitchellTJ 2004 Control of virulence by the two-component system CiaR/H is mediated via HtrA, a major virulence factor of Streptococcus pneumoniae. J Bacteriol 186 5258 5266

23. GuiralSMitchellTJMartinBClaverysJP 2005 Competence programmed predation of noncompetent cells in the human pathogen Streptococcus pneumoniae: genetic requirements. Proc Natl Acad Sci USA 102 8710 8715

24. ClaverysJPMartinBHåvarsteinLS 2007 Competence-induced fratricide in streptococci. Mol Microbiol 64 1423 1433

25. SungCKLiHClaverysJPMorrisonDA 2001 An rpsL Cassette, Janus, for Gene Replacement through Negative Selection in Streptococcus pneumoniae. Appl Environ Microbiol 67 5190 5196

26. SungCKMorrisonDA 2005 Two Distinct Functions of ComW in Stabilization and Activation of the Alternative Sigma Factor ComX in Streptococcus pneumoniae. J Bacteriol 187 3052 3061

27. MorrisonDABakerMF 1978 Competence for genetic transformation in pneumococcus depends on synthesis of a small set of proteins. Nature 282 215 217

28. GriffithF 1922 The significance of pneumococcal types. J Hygiene 27 113 159

29. OttolenghiEMacLeodCM 1963 Genetic transformation among living pneumococci in the mouse. Proc Natl Acad Sci U S A 50 417 419

30. ConantJESawyerWD 1967 Transformation during mixed pneumococcal infection of mice. J Bact 93 1869 1875

31. Auerbach-RubinFOttolenghi-NightingaleE 1971 Effect of eathanol on the clearance of airborne pneumococci and the rate of pneumococcal transformation in the lung. Infect Immun 3 688 693

32. OggioniMRIannelliFRicciSChiavoliniDParigiR 2004 Antibacterial activity of a competence-stimulating peptide in experimental sepsis caused by Streptococcus pneumoniae. Antimicrob Agents Chemother 48 4725 4732

33. CoomaraswamyG 1996 Induction of genetic transformation in Streptococcus pneumoniae by a pheromone peptide and its synthetic analogues. Ph.D. thesis Chicago University of Illinois

34. HavarsteinLGMorrisonDA 1999 Quorum sensing and peptide pheromones in streptococcal competence for genetic transformation. DunnyGMWinansSC Cell-Cell Signaling in Bacteria Washington, D.C. American Society for Microbiology 9 26

35. OggioniMRTrappettiCKadiogluACassoneMIannelliF 2006 Switch from planktonic to sessile life: a major event in pneumococcal pathogenesis. Mol Microbiol 61 1196 1210

36. LynchJP3rdZhanelGG 2009 Streptococcus pneumoniae: does antimicrobial resistance matter? Semin Respir Crit Care Med 30 210 238

37. JonesRNJacobsMRSaderHS 2010 Streptococcus pneumoniae resistance: implications for therapy of community-acquired bacterial pneumonia. Int J Antimicrob Agents 36 197 204

38. KlugmanKPLonksJR 2005 Hidden epidemic of macrolide-resistant pneumococci. Emerg Infect Dis 11 802 827

39. JacobsMR 2008 Antimicrobial-resistant Streptococcus pneumoniae: trends and management. Expert Rev Anti Infect Ther 6 619 635

40. PozziGIannelliFOggioniMRSantagatiMStefaniS 2004 Genetic elements carrying macrolide efflux genes in streptococci. Curr Drug Targets Infect Disord 4 203 206

41. WiddowsonCAKlugmanKP 1998 The molecular mechanisms of tetracycline resistance in the pneumococcus. Microb Drug Resist 4 79 84

42. CroucherNJWalkerDRomeroPLennardNPatersonGK 2009 Role of conjugative elements in the evolution of the multidrug-resistant pandemic clone Streptococcus pneumoniae Spain23F ST81. J Bacteriol 191 1480 1489

43. ShenXYangHYuSYaoKWangYYuanLYangY 2008 Macrolide-resistance mechanisms in Streptococcus pneumoniae isolates from Chinese children in association with genes of tetM and integrase of conjugative transposons 1545. Microb Drug Resist 14 155 161

44. DowsonCGHutchisonABranniganJAGeorgeRCHansmanD 1989 Horizontal transfer of penicillin-binding protein genes in penicillin-resistant clinical isolates of Streptococcus pneumoniae. Proc Nati Acad Sci USA 86 8842 8846

45. PrudhommeMAttaiechLSanchezGMartinBClaverysJP 2006 Antibiotic stress induces genetic transformability in the human pathogen Streptococcus pneumoniae. Science 313 89 92

46. Ottolenghi-NightingaleE 1969 Spontaneously occurring bacterial transformations in mice. J Bacteriol 100 445 452

47. Ottolenghi-NightingaleE 1972 Competence of pneumococcal isolates and bacterial transformations in man. Infect Immun 6 785 92

48. DunnyGMLeonardBA 1997 Cell-cell communication in gram-positive bacteria. Annu Rev Microbiol 51 527 564

49. KleerebezemMQuadriLEKuipersOPde VosWM 1997 Quorum sensing by peptide pheromones and two-component signal-transduction systems in Gram-positive bacteria. Mol Microbiol 24 895 904

50. SturmeMHKleerebezemMNakayamaJAkkermansADVaughaEE 2002 Cell to cell communication by autoinducing peptides in gram-positive bacteria. Antonie Van Leeuwenhoek 81 233 243

51. LyonGJNovickRP 2004 Peptide signaling in Staphylococcus aureus and other Gram-positive bacteria. Peptides 25 1389 1403

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

53. HåvarsteinLSHakenbeckRGaustadP 1997 Natural competence in the genus Streptococcus: evidence that streptococci can change pherotype by interspecies recombinational exchanges. J Bacteriol 179 6589 6594

54. KilianMPoulsenKBlomqvistTHåvarsteinLSBek-ThomsenM 2008 Evolution of Streptococcus pneumoniae and its close commensal relatives. PLoS One 3 e2683

55. AveryOTMacLeodCMMcCartyM 1944 Studies on the chemical nature of the substance inducing transformation of pneumococcal types: inductions of transformation by a desoxyribonucleic acid fraction isolated from pneumococcus type III. J Exp Med 79 137 158

56. OttolenghiEHotchkissRD 1960 Appearance of genetic transforming activity in pneumococcal cultures. Science 132 1257 1258

57. TettelinHNelsonKEPaulsenITEisenJAReadTD 2001 Complete genome sequence of a virulent isolate of Streptococcus pneumoniae. Science 293 498 506

58. KausmallyLJohnsborgOLundeMKnutsenEHåvarsteinLS 2005 Choline-binding protein D (CbpD) in Streptococcus pneumoniae is essential for competence-induced cell lysis. J Bacteriol 187 4338 4345

59. ClaverysJ-PDintilhacAPestovaEVMartinBMorrisonDA 1995 Construction and evaluation of new drug-resistance cassettes for gene disruption mutagenesis iin Streptococcus pneumoniae, using an ami test platform. Gene 164 123 128

60. EldholmVJohnsborgOStraumeDOhnstadHSBergKH 2010 CbpD is a murein hydrolase that requires a dual cell envelope binding specificity to kill target cells during fratricide. Mol Microbiol 76 905 917