The ArlRS Two-Component System Is a Novel Regulator of Agglutination and Pathogenesis

English version České info

Staphylococcus aureus is a prominent bacterial pathogen that is known to agglutinate in the presence of human plasma to form stable clumps. There is increasing evidence that agglutination aids S. aureus pathogenesis, but the mechanisms of this process remain to be fully elucidated. To better define this process, we developed both tube based and flow cytometry methods to monitor clumping in the presence of extracellular matrix proteins. We discovered that the ArlRS two-component system regulates the agglutination mechanism during exposure to human plasma or fibrinogen. Using divergent S. aureus strains, we demonstrated that arlRS mutants are unable to agglutinate, and this phenotype can be complemented. We found that the ebh gene, encoding the Giant Staphylococcal Surface Protein (GSSP), was up-regulated in an arlRS mutant. By introducing an ebh complete deletion into an arlRS mutant, agglutination was restored. To assess whether GSSP is the primary effector, a constitutive promoter was inserted upstream of the ebh gene on the chromosome in a wildtype strain, which prevented clump formation and demonstrated that GSSP has a negative impact on the agglutination mechanism. Due to the parallels of agglutination with infective endocarditis development, we assessed the phenotype of an arlRS mutant in a rabbit combined model of sepsis and endocarditis. In this model the arlRS mutant displayed a large defect in vegetation formation and pathogenesis, and this phenotype was partially restored by removing GSSP. Altogether, we have discovered that the ArlRS system controls a novel mechanism through which S. aureus regulates agglutination and pathogenesis.

Vyšlo v časopise: The ArlRS Two-Component System Is a Novel Regulator of Agglutination and Pathogenesis. PLoS Pathog 9(12): e32767. doi:10.1371/journal.ppat.1003819
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1003819

Souhrn

Zdroje

1. LowyFD (1998) Staphylococcus aureus infections. N Engl J Med 339: 520–532.

2. DeLeoFR, ChambersHF (2009) Reemergence of antibiotic-resistant Staphylococcus aureus in the genomics era. J Clin Invest 119: 2464–2474.

3. ChambersHF, DeleoFR (2009) Waves of resistance: Staphylococcus aureus in the antibiotic era. Nat Rev Microbiol 7: 629–641.

4. SomervilleGA, ProctorRA (2009) At the crossroads of bacterial metabolism and virulence factor synthesis in Staphylococci. Microbiol Mol Biol Rev 73: 233–248.

5. ParsekMR, SinghPK (2003) Bacterial biofilms: an emerging link to disease pathogenesis. Ann Rev Microbiol 57: 677–701.

6. DeleoFR, OttoM, KreiswirthBN, ChambersHF (2010) Community-associated meticillin-resistant Staphylococcus aureus. Lancet 375: 1557–1568.

7. BurmolleM, ThomsenTR, FazliM, DigeI, ChristensenL, et al. (2010) Biofilms in chronic infections - a matter of opportunity - monospecies biofilms in multispecies infections. FEMS Immunol Med Microbiol 59: 324–336.

8. KiedrowskiMR, HorswillAR (2011) New approaches for treating staphylococcal biofilm infections. Ann N Y Acad Sci 1241: 104–121.

9. FazliM, BjarnsholtT, Kirketerp-MollerK, JorgensenB, AndersenAS, et al. (2009) Nonrandom distribution of Pseudomonas aeruginosa and Staphylococcus aureus in chronic wounds. J Clin Microbiol 47: 4084–4089.

10. AlhedeM, KraghKN, QvortrupK, Allesen-HolmM, van GennipM, et al. (2011) Phenotypes of non-attached Pseudomonas aeruginosa aggregates resemble surface attached biofilm. PLoS One 6: e27943.

11. BjarnsholtT, JensenPO, FiandacaMJ, PedersenJ, HansenCR, et al. (2009) Pseudomonas aeruginosa biofilms in the respiratory tract of cystic fibrosis patients. Pediatr Pulmonol 44: 547–558.

12. HaaberJ, CohnMT, FreesD, AndersenTJ, IngmerH (2012) Planktonic aggregates of Staphylococcus aureus protect against common antibiotics. PLoS One 7: e41075.

13. McAdowM, KimHK, DedentAC, HendrickxAP, SchneewindO, et al. (2011) Preventing Staphylococcus aureus sepsis through the inhibition of its agglutination in blood. PLoS Pathog 7: e1002307.

14. ChengAG, McAdowM, KimHK, BaeT, MissiakasDM, et al. (2010) Contribution of coagulases towards Staphylococcus aureus disease and protective immunity. PLoS Pathog 6: e1001036.

15. FournierB, HooperDC (2000) A new two-component regulatory system involved in adhesion, autolysis, and extracellular proteolytic activity of Staphylococcus aureus. J Bacteriol 182: 3955–3964.

16. MemmiG, NairDR, CheungA (2012) Role of ArlRS in autolysis in methicillin-sensitive and methicillin-resistant Staphylococcus aureus strains. J Bacteriol 194: 759–767.

17. FournierB, KlierA, RapoportG (2001) The two-component system ArlS-ArlR is a regulator of virulence gene expression in Staphylococcus aureus. Mol Microbiol 41: 247–261.

18. ChristnerM, FrankeGC, SchommerNN, WendtU, WegertK, et al. (2010) The giant extracellular matrix-binding protein of Staphylococcus epidermidis mediates biofilm accumulation and attachment to fibronectin. Mol Microbiol 75: 187–207.

19. ClarkeSR, HarrisLG, RichardsRG, FosterSJ (2002) Analysis of Ebh, a 1.1-megadalton cell wall-associated fibronectin-binding protein of Staphylococcus aureus. Infect Immun 70: 6680–6687.

20. DuthieES (1955) The action of fibrinogen on certain pathogenic cocci. J Gen Microbiol 13: 383–393.

21. McAdowM, MissiakasDM, SchneewindO (2012) Staphylococcus aureus secretes coagulase and von Willebrand factor binding protein to modify the coagulation cascade and establish host infections. J Innate Immun 4: 141–148.

22. McDevittD, FrancoisP, VaudauxP, FosterTJ (1994) Molecular characterization of the clumping factor (fibrinogen receptor) of Staphylococcus aureus. Mol Microbiol 11: 237–248.

23. KimHK, KimHY, SchneewindO, MissiakasD (2011) Identifying protective antigens of Staphylococcus aureus, a pathogen that suppresses host immune responses. FASEB J 25: 3605–3612.

24. MazmanianSK, LiuG, Ton-ThatH, SchneewindO (1999) Staphylococcus aureus sortase, an enzyme that anchors surface proteins to the cell wall. Science 285: 760–763.

25. AbrahamNM, JeffersonKK (2012) Staphylococcus aureus clumping factor B mediates biofilm formation in the absence of calcium. Microbiology 158: 1504–1512.

26. O'NeillE, PozziC, HoustonP, HumphreysH, RobinsonDA, et al. (2008) A novel Staphylococcus aureus biofilm phenotype mediated by the fibronectin-binding proteins, FnBPA and FnBPB. J Bacteriol 190: 3835–3850.

27. MerinoN, Toledo-AranaA, Vergara-IrigarayM, ValleJ, SolanoC, et al. (2009) Protein A-mediated multicellular behavior in Staphylococcus aureus. J Bacteriol 191: 832–843.

28. BeenkenKE, BlevinsJS, SmeltzerMS (2003) Mutation of sarA in Staphylococcus aureus limits biofilm formation. Infect Immun 71: 4206–4211.

29. LauderdaleKJ, BolesBR, CheungAL, HorswillAR (2009) Interconnections between Sigma B, agr, and proteolytic activity in Staphylococcus aureus biofilm maturation. Infect Immun 77: 1623–1635.

30. BolesBR, HorswillAR (2008) Agr-mediated dispersal of Staphylococcus aureus biofilms. PLoS Pathog 4: e1000052.

31. KiedrowskiMR, KavanaughJS, MaloneCL, MootzJM, VoyichJM, et al. (2011) Nuclease modulates biofilm formation in community-associated methicillin-resistant Staphylococcus aureus. PLoS One 6: e26714.

32. Toledo-AranaA, MerinoN, Vergara-IrigarayM, DebarbouilleM, PenadesJR, et al. (2005) Staphylococcus aureus develops an alternative, ica-independent biofilm in the absence of the arlRS two-component system. J Bacteriol 187: 5318–5329.

33. LiangX, ZhengL, LandwehrC, LunsfordD, HolmesD, et al. (2005) Global regulation of gene expression by ArlRS, a two-component signal transduction regulatory system of Staphylococcus aureus. J Bacteriol 187: 5486–5492.

34. MolkanenT, TyynelaJ, HelinJ, KalkkinenN, KuuselaP (2002) Enhanced activation of bound plasminogen on Staphylococcus aureus by staphylokinase. FEBS Lett 517: 72–78.

35. ChristnerRB, BoyleMD (1996) Role of staphylokinase in the acquisition of plasmin(ogen)-dependent enzymatic activity by staphylococci. J Infect Dis 173: 104–112.

36. SakamotoS, TanakaY, TanakaI, TakeiT, YuJ, et al. (2008) Electron microscopy and computational studies of Ebh, a giant cell-wall-associated protein from Staphylococcus aureus. Biochem Biophys Res Commun 376: 261–266.

37. TanakaY, SakamotoS, KurodaM, GodaS, GaoYG, et al. (2008) A helical string of alternately connected three-helix bundles for the cell wall-associated adhesion protein Ebh from Staphylococcus aureus. Structure 16: 488–496.

38. FeyPD, EndresJL, YajjalaVK, WidhelmTJ, BoissyRJ, et al. (2013) A genetic resource for rapid and comprehensive phenotype screening of nonessential Staphylococcus aureus genes. MBio 4: e00537–00512.

39. Bubeck WardenburgJ, SchneewindO (2008) Vaccine protection against Staphylococcus aureus pneumonia. J Exp Med 205: 287–294.

40. KobayashiSD, MalachowaN, WhitneyAR, BraughtonKR, GardnerDJ, et al. (2011) Comparative analysis of USA300 virulence determinants in a rabbit model of skin and soft tissue infection. J Infect Dis 204: 937–941.

41. ThoendelM, KavanaughJS, FlackCE, HorswillAR (2011) Peptide signaling in the staphylococci. Chem Rev 111: 117–151.

42. BentonBM, ZhangJP, BondS, PopeC, ChristianT, et al. (2004) Large-scale identification of genes required for full virulence of Staphylococcus aureus. J Bacteriol 186: 8478–8489.

43. SchlievertPM (2009) Cytolysins, superantigens, and pneumonia due to community-associated methicillin-resistant Staphylococcus aureus. J Infect Dis 200: 676–678.

44. SchlievertPM, Chuang-SmithON, PetersonML, CookLC, DunnyGM (2010) Enterococcus faecalis endocarditis severity in rabbits is reduced by IgG Fabs interfering with aggregation substance. PLoS One 5: e13194.

45. SpauldingAR, SatterwhiteEA, LinYC, Chuang-SmithON, FrankKL, et al. (2012) Comparison of Staphylococcus aureus strains for ability to cause infective endocarditis and lethal sepsis in rabbits. Frontiers in cellular and infection microbiology 2: 18.

46. BeenkenKE, MrakLN, GriffinLM, ZielinskaAK, ShawLN, et al. (2010) Epistatic relationships between sarA and agr in Staphylococcus aureus biofilm formation. PLoS One 5: e10790.

47. ZielinskaAK, BeenkenKE, MrakLN, SpencerHJ, PostGR, et al. (2012) sarA-mediated repression of protease production plays a key role in the pathogenesis of Staphylococcus aureus USA300 isolates. Molecular microbiology 86: 1183–1196.

48. WalkerJN, HorswillAR (2012) A coverslip-based technique for evaluating Staphylococcus aureus biofilm formation on human plasma. Front Cell Infect Microbiol 2: 39.

49. RothforkJM, Dessus-BabusS, Van WamelWJ, CheungAL, GreshamHD (2003) Fibrinogen depletion attenuates Staphyloccocus aureus infection by preventing density-dependent virulence gene up-regulation. J Immunol 171: 5389–5395.

50. KapralFA (1966) Clumping of Staphylococcus aureus in the peritoneal cavity of mice. J Bacteriol 92: 1188–1195.

51. ChengAG, DeDentAC, SchneewindO, MissiakasD (2011) A play in four acts: Staphylococcus aureus abscess formation. Trends Microbiol 19: 225–232.

52. KapralFA, GodwinJR, DyeES (1980) Formation of intraperitoneal abscesses by Staphylococcus aureus. Infect Immun 30: 204–211.

53. KurodaM, TanakaY, AokiR, ShuD, TsumotoK, et al. (2008) Staphylococcus aureus giant protein Ebh is involved in tolerance to transient hyperosmotic pressure. Biochem Biophys Res Commun 374: 237–241.

54. MoreillonP, QueYA (2004) Infective endocarditis. Lancet 363: 139–149.

55. NienaberJJ, Sharma KuinkelBK, Clarke-PearsonM, LamlertthonS, ParkL, et al. (2011) Methicillin-susceptible Staphylococcus aureus endocarditis isolates are associated with clonal complex 30 genotype and a distinct repertoire of enterotoxins and adhesins. J Infect Dis 204: 704–713.

56. McCarthyAJ, LindsayJA (2010) Genetic variation in Staphylococcus aureus surface and immune evasion genes is lineage associated: implications for vaccine design and host-pathogen interactions. BMC microbiology 10: 173.

57. SchenkS, LaddagaRA (1992) Improved method for electroporation of Staphylococcus aureus. FEMS Microbiol Lett 73: 133–138.

58. NovickRP (1991) Genetic systems in staphylococci. Methods Enzymol 204: 587–636.

59. WalterJ, SchwabC, LoachDM, GanzleMG, TannockGW (2008) Glucosyltransferase A (GtfA) and inulosucrase (Inu) of Lactobacillus reuteri TMW1.106 contribute to cell aggregation, in vitro biofilm formation, and colonization of the mouse gastrointestinal tract. Microbiology 154: 72–80.

60. WormannME, ReichmannNT, MaloneCL, HorswillAR, GrundlingA (2011) Proteolytic cleavage inactivates the Staphylococcus aureus lipoteichoic acid synthase. J Bacteriol 193: 5279–5291.

61. LauderdaleKJ, MaloneCL, BolesBR, MorcuendeJ, HorswillAR (2010) Biofilm dispersal of community-associated methicillin-resistant Staphylococcus aureus on orthopedic implant material. J Orthop Res 28: 55–61.

62. KolarSL, Antonio IbarraJ, RiveraFE, MootzJM, DavenportJE, et al. (2012) Extracellular proteases are key mediators of Staphylococcus aureus virulence via the global modulation of virulence-determinant stability. MicrobiologyOpen 2: 18–34.

63. PangYY, SchwartzJ, ThoendelM, AckermannLW, HorswillAR, et al. (2010) agr-Dependent interactions of Staphylococcus aureus USA300 with human polymorphonuclear neutrophils. J Innate Immun 2: 546–559.

64. NairD, MemmiG, HernandezD, BardJ, BeaumeM, et al. (2011) Whole-genome sequencing of Staphylococcus aureus strain RN4220, a key laboratory strain used in virulence research, identifies mutations that affect not only virulence factors but also the fitness of the strain. J Bacteriol 193: 2332–2335.

65. BolesBR, ThoendelM, RothAJ, HorswillAR (2010) Identification of genes involved in polysaccharide-independent Staphylococcus aureus biofilm formation. PLoS One 5: e10146.

66. VoyichJM, VuongC, DeWaldM, NygaardTK, KocianovaS, et al. (2009) The SaeR/S gene regulatory system is essential for innate immune evasion by Staphylococcus aureus. J Infect Dis 199: 1698–1706.

67. BabaT, TakeuchiF, KurodaM, YuzawaH, AokiK, et al. (2002) Genome and virulence determinants of high virulence community-acquired MRSA. Lancet 359: 1819–1827.

68. BabaT, BaeT, SchneewindO, TakeuchiF, HiramatsuK (2008) Genome sequence of Staphylococcus aureus strain Newman and comparative analysis of staphylococcal genomes: polymorphism and evolution of two major pathogenicity islands. J Bacteriol 190: 300–310.