RocA Truncation Underpins Hyper-Encapsulation, Carriage Longevity and Transmissibility of Serotype M18 Group A Streptococci

English version České info

Group A streptococcal isolates of serotype M18 are historically associated with epidemic waves of pharyngitis and the non-suppurative immune sequela rheumatic fever. The serotype is defined by a unique, highly encapsulated phenotype, yet the molecular basis for this unusual colony morphology is unknown. Here we identify a truncation in the regulatory protein RocA, unique to and conserved within our serotype M18 GAS collection, and demonstrate that it underlies the characteristic M18 capsule phenotype. Reciprocal allelic exchange mutagenesis of rocA between M18 GAS and M89 GAS demonstrated that truncation of RocA was both necessary and sufficient for hyper-encapsulation via up-regulation of both precursors required for hyaluronic acid synthesis. Although RocA was shown to positively enhance covR transcription, quantitative proteomics revealed RocA to be a metabolic regulator with activity beyond the CovR/S regulon. M18 GAS demonstrated a uniquely protuberant chain formation following culture on agar that was dependent on excess capsule and the RocA mutation. Correction of the M18 rocA mutation reduced GAS survival in human blood, and in vivo naso-pharyngeal carriage longevity in a murine model, with an associated drop in bacterial airborne transmission during infection. In summary, a naturally occurring truncation in a regulator explains the encapsulation phenotype, carriage longevity and transmissibility of M18 GAS, highlighting the close interrelation of metabolism, capsule and virulence.

Vyšlo v časopise: RocA Truncation Underpins Hyper-Encapsulation, Carriage Longevity and Transmissibility of Serotype M18 Group A Streptococci. PLoS Pathog 9(12): e32767. doi:10.1371/journal.ppat.1003842
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1003842

Souhrn

Zdroje

1. CourtneyHS, HastyDL (2002) DaleJB (2002) Molecular mechanisms of adhesion, colonization, and invasion of group A streptococci. Ann Med 34 : 77–87.

2. WesselsMR, MosesAE, GoldbergJB, DiCesareTJ (1991) Hyaluronic acid capsule is a virulence factor for mucoid group A streptococci. Proc Natl Acad Sci 88 : 8317–8321.

3. WesselsMR, BronzeMS (1994) Critical role of the group A streptococcal capsule in pharyngeal colonization and infection in mice. Proc Natl Acad Sci 91 : 12238–12242.

4. CywesC, StamenkovicI, WesselsMR (2000) CD44 as a receptor for colonization of the pharynx by group A Streptococcus. J Clin Invest 106 : 995–1002.

5. CywesC, WesselsMR (2001) Group A Streptococcus tissue invasion by CD44-mediated cell signaling. Nature 414 : 648–652.

6. SmootJC, KorgenskiEK, DalyJA, VeasyLG, MusserJM (2002) Molecular analysis of group A Streptococcus type emm18 isolates temporally associated with acute rheumatic fever outbreaks in Salt Lake City, Utah. J Clin Microbiol 40 : 1805–1810.

7. SmootJC, BarbianKD, Van GompelJJ, SmootLM, ChausseeMS, et al. (2002) Genome sequence and comparative microarray analysis of serotype M18 group A Streptococcus strains associated with acute rheumatic fever outbreaks. Proc Natl Acad Sci 99 : 4668–4673.

8. JohnsonD, StevensDL, KaplanEL (1992) Epidemiologic analysis of group A streptococcal serotypes associated with severe systemic infections, rheumatic fever, or uncomplicated pharyngitis. J Infect Dis 30 : 374.

9. VeasyLG, TaniLY, DalyJA, KorgenskiK, MineL, et al. (2004) Temporal association of the appearance of mucoid strains of Streptococcus pyogenes with a continuing high incidence of rheumatic fever in Utah. Pediatrics 113: e168–e172.

10. VeasyLG, WiedmeierSE, OrsmondGS, RuttenbergHD, BoucekMM, et al. (1987) Resurgence of acute rheumatic fever in the intermountain area of the United States. N Engl J Med 316 : 421–427.

11. EllisNM, KuraharaDK, VohraH, Mascaro-BlancoA, ErdemG, et al. (2010) Priming the immune system for heart disease: a perspective on group A streptococci. J Infect Dis 202 : 1059–1067.

12. AlbertiS, AshbaughCD, WesselsMR (1998) Structure of the has operon promoter and regulation of hyaluronic acid capsule expression in group A Streptococcus. Mol Microbiol 28 : 343–353.

13. GrahamMR, SmootLM, MigliaccioCA, VirtanevaK, SturdevantDE, et al. (2002) Virulence control in group A Streptococcus by a two-component gene regulatory system: global expression profiling and in vivo infection modeling. Proc Natl Acad Sci 99 : 13855–13860.

14. van der RijnI, DrakeRR (1992) Analysis of the streptococcal hyaluronic acid synthase complex using the photoaffinity probe 5-azido-UDP-glucuronic acid. J Biol Chem 267 : 24302–24306.

15. BlankLM, HugenholtzP, NielsenLK (2008) Evolution of the hyaluronic acid synthesis (has) operon in Streptococcus zooepidemicus and other pathogenic streptococci. J Mol Evol 67 : 13–22.

16. FloresAR, JewellBE, FittipaldiN, BeresSB, MusserJM (2012) Human disease isolates of serotype M4 and M22 group A streptococcus lack genes required for hyaluronic acid capsule biosynthesis. mBio 3(6): e00413–12.

17. ChoKH, CaparonMG (2005) Patterns of virulence gene expression differ between biofilm and tissue communities of Streptococcus pyogenes. Mol Microbiol 2005 Sep;57(6): 1545–56.

18. BrowningDF, BusbySJ (2004) The regulation of bacterial transcription initiation. Nat Rev Microbiol 2 : 57–65.

19. CaparonMG, GeistRT, Perez-CasalJ, ScottJR (1992) Environmental regulation of virulence in group A streptococci: transcription of the gene encoding M protein is stimulated by carbon dioxide. J Bacteriol 174 : 5693–5701.

20. GrahamMR, VirtanevaK, PorcellaSF, BarryWT, GowenBB, et al. (2005) Group A Streptococcus transcriptome dynamics during growth in human blood reveals bacterial adaptive and survival strategies. Am J Pathol 166 : 455–465.

21. HynesW (2004) Virulence factors of the group A streptococci and genes that regulate their expression. Front Biosci 9 : 3399–3433.

22. KreikemeyerB, McIverKS, PodbielskiA (2003) Virulence factor regulation and regulatory networks in Streptococcus pyogenes and their impact on pathogen-host interactions. Trends Microbiol 11 : 224–232.

23. PritchardKH, ClearyPP (1996) Differential expression of genes in the vir regulon of Streptococcus pyogenes is controlled by transcription termination. Mol Gen Genet 250 : 207–213.

24. SumbyP, WhitneyAR, GravissEA, DeLeoFR, MusserJM (2006) Genome-wide analysis of group A streptococci reveals a mutation that modulates global phenotype and disease specificity. PLoS Pathog 2: e5.

25. HeathA, DiRitaVJ, BargNL, EnglebergNC (1999) A two-component regulatory system, CsrR-CsrS, represses expression of three Streptococcus pyogenes virulence factors, hyaluronic acid capsule, streptolysin S, and pyrogenic exotoxin B. Infect Immun 67 : 5298–5305.

26. WalkerMJ, HollandsA, Sanderson-SmithML, ColeJN, KirkJK, et al. (2007) DNase Sda1 provides selection pressure for a switch to invasive group A streptococcal infection. Nat Med 13 : 981–985.

27. TurnerCE, KurupatiP, JonesMD, EdwardsRJ, SriskandanS (2009) Emerging role of the interleukin-8 cleaving enzyme SpyCEP in clinical Streptococcus pyogenes Infection. J Infect Dis 200 : 555–563.

28. BiswasI, ScottJR (2003) Identification of rocA, a positive regulator of covR expression in the group A streptococcus. J Bacteriol 185 : 3081–3090.

29. LevinJC, WesselsMR (1998) Identification of csrR/csrS, a genetic locus that regulates hyaluronic acid capsule synthesis in group A Streptococcus. Mol Microbiol 30 : 209–219.

30. TreviñoJ, PerezN, Ramirez-PeñaE, LiuZ, ShelburneSA3rd, et al. (2009) CovS simultaneously activates and inhibits the CovR-mediated repression of distinct subsets of group A Streptococcus virulence factor-encoding genes. Infect Immun 77(8): 3141–9.

31. EnglebergNC, HeathA, MillerA, RiveraC, DiRitaVJ (2001) Spontaneous mutations in the CsrRS two-component regulatory system of Streptococcus pyogenes result in enhanced virulence in a murine model of skin and soft tissue infection. J Infect Dis 183 : 1043–54.

32. HollandsA, PenceMA, TimmerAM, OsvathSR, TurnbullL, et al. (2010) Genetic switch to hypervirulence reduces colonization phenotypes of the globally disseminated group A streptococcus M1T1 clone. J Infect Dis 202(1): 11–9.

33. BarnettTC, BugryshevaJV, ScottJR (2007) Role of mRNA stability in growth phase regulation of gene expression in the group A streptococcus. J Bacteriol 189(5): 1866–73.

34. ShelburneSAIII, SumbyP, SitkiewiczI, GranvilleC, DeLeoFR, et al. (2005) Central role of a bacterial two-component gene regulatory system of previously unknown function in pathogen persistence in human saliva. Proc Natl Acad Sci 102 : 16037–16042.

35. SumbyP, ZhangS, WhitneyAR, FalugiF, GrandiG, et al. (2008) A chemokine-degrading extracellular protease made by group A Streptococcus alters pathogenesis by enhancing evasion of the innate immune response. Infect Immun 76 : 978–985.

36. SugarevaV, ArltR, FiedlerT, RianiC, PodbielskiA, et al. (2010) Serotype -⁠ and strain -⁠ dependent contribution of the sensor kinase CovS of the CovRS two-component system to Streptococcus pyogenes pathogenesis. BMC Microbiol 10 : 34.

37. ChowV, NongG, PrestonJF (2007) Structure, function, and regulation of the aldouronate utilization gene cluster from Paenibacillus sp. strain JDR-2. J J Bacteriol 189 : 8863–8870.

38. DinklaK, RohdeM, JansenWT, KaplanEL, ChhatwalGS, et al. (2003) Rheumatic fever-associated Streptococcus pyogenes isolates aggregate collagen. J Clin Invest 111(12): 1905–12.

39. CortésG, WesselsMR (2007) Inhibition of dendritic cell maturation by group A Streptococcus. J Infect Dis 2009 Oct 1;200(7): 1152–61.

40. TermeerC, BenedixF, SleemanJ, FieberC, VoithU, et al. (2002) Oligosaccharides of Hyaluronan activate dendritic cells via toll-like receptor 4. J Exp Med 7;195(1): 99–111.

41. HollandsA, AzizRK, KansalR, KotbM, NizetV, et al. (2008) A naturally occurring mutation in ropB suppresses SpeB expression and reduces M1T1 group A streptococcal systemic virulence. PLoS One 3(12): e4102.

42. OlsenRJ, SitkiewiczI, AyerasAA, GonulalVE, CantuC, et al. (2010) Decreased necrotizing fasciitis capacity caused by a single nucleotide mutation that alters a multiple gene virulence axis. Proc Natl Acad Sci U S A 12;107(2): 888–93.

43. AlamFM, TurnerCE, SmithK, WilesS, SriskandanS (2013) Inactivation of the CovR/S virulence regulator impairs infection in an improved murine model of Streptococcus pyogenes naso-pharyngeal infection. PLoS ONE 8: e61655.

44. PospiechA, NeumannB (1995) A versatile quick-prep of genomic DNA from gram-positive bacteria. Trends Genet 11 : 217–218.

45. UnnikrishnanM, CohenJ, SriskandanS (2001) Complementation of a speA negative Streptococcus pyogenes with speA: effects on virulence and production of streptococcal pyrogenic exotoxin A. Microb Pathog 31 : 109–114.

46. SriskandanS, UnnikrishnanM, KrauszT, CohenJ (2000) Mitogenic factor (MF) is the major DNase of serotype M89 Streptococcus pyogenes. Microbiology 146(Pt 11): 2785–92.

47. EdwardsRJ, WrigleyA, BaiZ, BatemanM, RussellH, et al. (2007) C-terminal antibodies (CTAbs): a simple and broadly applicable approach for the rapid generation of protein-specific antibodies with predefined specificity. Proteomics 7 : 1364–1372.

48. ZinkernagelAS, TimmerAM, PenceMA, LockeJB, BuchananJT, et al. (2008) The IL-8 protease SpyCEP/ScpC of group A Streptococcus promotes resistance to neutrophil killing. Cell Host Microbe 14;4(2): 170–8.

49. MalickLE, WilsonRB (1975) Modified thiocarbohydrazide procedure for scanning electron microscopy: routine use for normal, pathological, or experimental tissues. Stain Technol 50 : 265–269.