A Combination of Independent Transcriptional Regulators Shapes Bacterial Virulence Gene Expression during Infection


Transcriptional regulatory networks are fundamental to how microbes alter gene expression in response to environmental stimuli, thereby playing a critical role in bacterial pathogenesis. However, understanding how bacterial transcriptional regulatory networks function during host-pathogen interaction is limited. Recent studies in group A Streptococcus (GAS) suggested that the transcriptional regulator catabolite control protein A (CcpA) influences many of the same genes as the control of virulence (CovRS) two-component gene regulatory system. To provide new information about the CcpA and CovRS networks, we compared the CcpA and CovR transcriptomes in a serotype M1 GAS strain. The transcript levels of several of the same genes encoding virulence factors and proteins involved in basic metabolic processes were affected in both ΔccpA and ΔcovR isogenic mutant strains. Recombinant CcpA and CovR bound with high-affinity to the promoter regions of several co-regulated genes, including those encoding proteins involved in carbohydrate and amino acid metabolism. Compared to the wild-type parental strain, ΔccpA and ΔcovRΔccpA isogenic mutant strains were significantly less virulent in a mouse myositis model. Inactivation of CcpA and CovR alone and in combination led to significant alterations in the transcript levels of several key GAS virulence factor encoding genes during infection. Importantly, the transcript level alterations in the ΔccpA and ΔcovRΔccpA isogenic mutant strains observed during infection were distinct from those occurring during growth in laboratory medium. These data provide new knowledge regarding the molecular mechanisms by which pathogenic bacteria respond to environmental signals to regulate virulence factor production and basic metabolic processes during infection.


Vyšlo v časopise: A Combination of Independent Transcriptional Regulators Shapes Bacterial Virulence Gene Expression during Infection. PLoS Pathog 6(3): e32767. doi:10.1371/journal.ppat.1000817
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1000817

Souhrn

Transcriptional regulatory networks are fundamental to how microbes alter gene expression in response to environmental stimuli, thereby playing a critical role in bacterial pathogenesis. However, understanding how bacterial transcriptional regulatory networks function during host-pathogen interaction is limited. Recent studies in group A Streptococcus (GAS) suggested that the transcriptional regulator catabolite control protein A (CcpA) influences many of the same genes as the control of virulence (CovRS) two-component gene regulatory system. To provide new information about the CcpA and CovRS networks, we compared the CcpA and CovR transcriptomes in a serotype M1 GAS strain. The transcript levels of several of the same genes encoding virulence factors and proteins involved in basic metabolic processes were affected in both ΔccpA and ΔcovR isogenic mutant strains. Recombinant CcpA and CovR bound with high-affinity to the promoter regions of several co-regulated genes, including those encoding proteins involved in carbohydrate and amino acid metabolism. Compared to the wild-type parental strain, ΔccpA and ΔcovRΔccpA isogenic mutant strains were significantly less virulent in a mouse myositis model. Inactivation of CcpA and CovR alone and in combination led to significant alterations in the transcript levels of several key GAS virulence factor encoding genes during infection. Importantly, the transcript level alterations in the ΔccpA and ΔcovRΔccpA isogenic mutant strains observed during infection were distinct from those occurring during growth in laboratory medium. These data provide new knowledge regarding the molecular mechanisms by which pathogenic bacteria respond to environmental signals to regulate virulence factor production and basic metabolic processes during infection.


Zdroje

1. SmithH

1958 The use of bacteria grown in vivo for studies on the basis of their pathogenicity. Annu Rev Microbiol 12 77 102

2. Toledo-AranaA

DussurgetO

NikitasG

SestoN

Guet-RevilletH

2009 The Listeria transcriptional landscape from saprophytism to virulence. Nature 459 950 956

3. HungDT

ShakhnovichEA

PiersonE

MekalanosJJ

2005 Small-molecule inhibitor of Vibrio cholerae virulence and intestinal colonization. Science 310 670 674

4. CheungAL

BayerAS

ZhangG

GreshamH

XiongYQ

2004 Regulation of virulence determinants in vitro and in vivo in Staphylococcus aureus. FEMS Immunol Med Microbiol 40 1 9

5. YoonH

McDermottJE

PorwollikS

McClellandM

HeffronF

2009 Coordinated regulation of virulence during systemic infection of Salmonella enterica serovar Typhimurium. PLoS Pathog 5 e1000306 doi:10.1371/journal.ppat.1000306

6. TartAH

WalkerMJ

MusserJM

2007 New understanding of the group A Streptococcus pathogenesis cycle. Trends Microbiol 15 318 325

7. ChurchwardG

2007 The two faces of Janus: virulence gene regulation by CovR/S in group A streptococci. Mol Microbiol 64 34 41

8. OlsenRJ

ShelburneSA

MusserJM

2009 Molecular mechanisms underlying group A streptococcal pathogenesis. Cell Microbiol 11 1 12

9. HondorpER

McIverKS

2007 The Mga virulence regulon: infection where the grass is greener. Mol Microbiol 66 1056 1065

10. HorsburghMJ

AishJL

WhiteIJ

ShawL

LithgowJK

2002 sigmaB modulates virulence determinant expression and stress resistance: characterization of a functional rsbU strain derived from Staphylococcus aureus 8325-4. J Bacteriol 184 5457 5467

11. KazmierczakMJ

WiedmannM

BoorKJ

2005 Alternative sigma factors and their roles in bacterial virulence. Microbiol Mol Biol Rev 69 527 543

12. 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

13. McIverKS

2009 Stand-alone response regulators controlling global virulence networks in streptococcus pyogenes. Contrib Microbiol 16 103 119

14. StockAM

RobinsonVL

GoudreauPN

2000 Two-component signal transduction. Annu Rev Biochem 69 183 215

15. GaoR

MackTR

StockAM

2007 Bacterial response regulators: versatile regulatory strategies from common domains. Trends Biochem Sci 32 225 234

16. TrevinoJ

PerezN

Ramirez-PenaE

LiuZ

ShelburneSA3rd

2009 CovS simultaneously activates and inhibits the CovR-mediated repression of distinct subsets of group A Streptococcus virulence factor-encoding genes. Infect Immun 77 3141 3149

17. 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

18. MillerAA

EnglebergNC

DiRitaVJ

2001 Repression of virulence genes by phosphorylation-dependent oligomerization of CsrR at target promoters in Streptococcus pyogenes. Mol Microbiol 40 976 990

19. FederleMJ

McIverKS

ScottJR

1999 A response regulator that represses transcription of several virulence operons in the group A Streptococcus. J Bacteriol 181 3649 3657

20. GrahamMR

SmootLM

MigliaccioCA

VirtanevaK

SturdevantDE

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 USA 99 13855 13860

21. 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 1054

22. GrahamMR

VirtanevaK

PorcellaSF

GardnerDJ

LongRD

2006 Analysis of the transcriptome of group A Streptococcus in mouse soft tissue infection. Am J Pathol 169 927 942

23. DaltonTL

ScottJR

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

24. GaoJ

GusaAA

ScottJR

ChurchwardG

2005 Binding of the global response regulator protein CovR to the sag promoter of Streptococcus pyogenes reveals a new mode of CovR-DNA interaction. J Biol Chem 280 38948 38956

25. FederleMJ

ScottJR

2002 Identification of binding sites for the group A streptococcal global regulator CovR. Mol Microbiol 43 1161 1172

26. 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 doi:10.1371/journal.ppat.0020005

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. WalkerMJ

HollandsA

Sanderson-SmithML

ColeJN

KirkJK

2007 DNase Sda1 provides selection pressure for a switch to invasive group A streptococcal infection. Nat Med 13 981 985

29. DaltonTL

CollinsJT

BarnettTC

ScottJR

2006 RscA, a member of the MDR1 family of transporters, is repressed by CovR and required for growth of Streptococcus pyogenes under heat stress. J Bacteriol 188 77 85

30. ShelburneSA3rd

KeithD

HorstmannN

SumbyP

DavenportMT

2008 A direct link between carbohydrate utilization and virulence in the major human pathogen group A Streptococcus. Proc Natl Acad Sci USA 105 1698 1703

31. KinkelTL

McIverKS

2008 CcpA-mediated repression of streptolysin S expression and virulence in the group A Streptococcus. Infect Immun 76 3451 3463

32. IyerR

BaligaNS

CamilliA

2005 Catabolite control protein A (CcpA) contributes to virulence and regulation of sugar metabolism in Streptococcus pneumoniae. J Bacteriol 187 8340 8349

33. SeidlK

BischoffM

Berger-BachiB

2008 CcpA mediates the catabolite repression of tst in Staphylococcus aureus. Infect Immun 76 5093 5099

34. AbranchesJ

NascimentoMM

ZengL

BrowngardtCM

WenZT

2008 CcpA regulates central metabolism and virulence gene expression in Streptococcus mutans. J Bacteriol 190 2340 2349

35. VargaJ

StirewaltVL

MelvilleSB

2004 The CcpA protein is necessary for efficient sporulation and enterotoxin gene (cpe) regulation in Clostridium perfringens. J Bacteriol 186 5221 5229

36. SonensheinAL

2007 Control of key metabolic intersections in Bacillus subtilis. Nat Rev Microbiol 5 917 927

37. SchumacherMA

AllenGS

DielM

SeidelG

HillenW

2004 Structural basis for allosteric control of the transcription regulator CcpA by the phosphoprotein HPr-Ser46-P. Cell 118 731 741

38. DeutscherJ

FranckeC

PostmaPW

2006 How phosphotransferase system-related protein phosphorylation regulates carbohydrate metabolism in bacteria. Microbiol Mol Biol Rev 70 939 1031

39. KietzmanCC

CaparonMG

2010 CcpA and LacD.1 affect temporal regulation of S. pyogenes virulence genes. Infect Immun 78 241 52

40. LiangX

ZhengL

LandwehrC

LunsfordD

HolmesD

2005 Global regulation of gene expression by ArlRS, a two-component signal transduction regulatory system of Staphylococcus aureus. J Bacteriol 187 5486 5492

41. SeidlK

MullerS

FrancoisP

KriebitzschC

SchrenzelJ

2009 Effect of a glucose impulse on the CcpA regulon in Staphylococcus aureus. BMC Microbiol 9 95

42. BiswasS

BiswasI

2006 Regulation of the glucosyltransferase (gtfBC) operon by CovR in Streptococcus mutans. J Bacteriol 188 988 998

43. SomervilleGA

ProctorRA

2009 At the crossroads of bacterial metabolism and virulence factor synthesis in staphylococci. Microbiol Mol Biol Rev 73 233 248

44. LukomskiS

SreevatsanS

AmbergC

ReichardtW

WoischnikM

1997 Inactivation of Streptococcus pyogenes extracellular cysteine protease significantly decreases mouse lethality of serotype M3 and M49 strains. J Clin Invest 99 2574 2580

45. HakanssonA

BentleyCC

ShakhnovicEA

WesselsMR

2005 Cytolysin-dependent evasion of lysosomal killing. Proc Natl Acad Sci USA 102 5192 5197

46. SumbyP

ZhangS

WhitneyAR

FalugiF

GrandiG

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

47. CollinM

OlsenA

2001 EndoS, a novel secreted protein from Streptococcus pyogenes with endoglycosidase activity on human IgG. EMBO J 20 3046 3055

48. AndersonBJ

LarkinC

GujaK

SchildbachJF

2008 Using fluorophore-labeled oligonucleotides to measure affinities of protein-DNA interactions. Methods Enzymol 450 253 272

49. BernishB

van de RijnI

1999 Characterization of a two-component system in Streptococcus pyogenes which is involved in regulation of hyaluronic acid production. J Biol Chem 274 4786 4793

50. ShelburneSA3rd

SumbyP

SitkiewiczI

GranvilleC

DeLeoFR

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 USA 102 16037 16042

51. GryllosI

Tran-WinklerHJ

ChengMF

ChungH

BolcomeR3rd

2008 Induction of group A Streptococcus virulence by a human antimicrobial peptide. Proc Natl Acad Sci USA 105 16755 16760

52. TurnerCE

KurupatiP

WilesS

EdwardsRJ

SriskandanS

2009 Impact of immunization against SpyCEP during invasive disease with two streptococcal species: Streptococcus pyogenes and Streptococcus equi. Vaccine 27 4923 4929

53. JacobF

MonodJ

1961 Genetic regulatory mechanisms in the synthesis of proteins. J Mol Biol 3 318 356

54. CallahanLT3rd

RichardsonSH

1973 Biochemistry of Vibrio cholerae virulence. 3. Nutritional requirements for toxin production and the effects of pH on toxin elaboration in chemically defined media. Infect Immun 7 567 572

55. RegassaLB

NovickRP

BetleyMJ

1992 Glucose and nonmaintained pH decrease expression of the accessory gene regulator (agr) in Staphylococcus aureus. Infect Immun 60 3381 3388

56. GryllosI

GrifantiniR

ColapricoA

JiangS

DeforceE

2007 Mg(2+) signaling defines the group A streptococcal CsrRS (CovRS) regulon. Mol Microbiol 65 671 683

57. XiongYQ

WillardJ

YeamanMR

CheungAL

BayerAS

2006 Regulation of Staphylococcus aureus alpha-toxin gene (hla) expression by agr, sarA, and sae in vitro and in experimental infective endocarditis. J Infect Dis 194 1267 1275

58. FroehlichBJ

BatesC

ScottJR

2009 Streptococcus pyogenes CovRS mediates growth in iron starvation and in the presence of the human cationic antimicrobial peptide LL-37. J Bacteriol 191 673 677

59. MiwaY

SaikawaM

FujitaY

1994 Possible function and some properties of the CcpA protein of Bacillus subtilis. Microbiology 140 2567 2575

60. LorcaGL

ChungYJ

BaraboteRD

WeylerW

SchillingCH

2005 Catabolite repression and activation in Bacillus subtilis: dependency on CcpA, HPr, and HprK. J Bacteriol 187 7826 7839

61. MittalS

KroosL

2009 A combination of unusual transcription factors binds cooperatively to control Myxococcus xanthus developmental gene expression. Proc Natl Acad Sci USA 106 1965 1970

62. RaskoDA

MoreiraCG

Li deR

ReadingNC

RitchieJM

2008 Targeting QseC signaling and virulence for antibiotic development. Science 321 1078 1080

63. ChaffinDO

RubensCE

1998 Blue/white screening of recombinant plasmids in Gram-positive bacteria by interruption of alkaline phosphatase gene (phoZ) expression. Gene 219 91 99

64. VirtanevaK

PorcellaSF

GrahamMR

IrelandRM

JohnsonCA

2005 Longitudinal analysis of the group A Streptococcus transcriptome in experimental pharyngitis in cynomolgus macaques. Proc Natl Acad Sci USA 102 9014 9019

65. KapurV

TopouzisS

MajeskyMW

LiLL

HamrickMR

1993 A conserved Streptococcus pyogenes extracellular cysteine protease cleaves human fibronectin and degrades vitronectin. Microb Pathog 15 327 346

66. GusaAA

GaoJ

StringerV

ChurchwardG

ScottJR

2006 Phosphorylation of the group A streptococcal CovR response regulator causes dimerization and promoter-specific recruitment by RNA polymerase. J Bacteriol 188 4620 4626

67. SumbyP

PorcellaSF

MadrigalAG

BarbianKD

VirtanevaK

2005 Evolutionary origin and emergence of a highly successful clone of serotype M1 group A Streptococcus involved multiple horizontal gene transfer events. J Infect Dis 192 771 782

68. LukomskiS

HoeNP

AbdiI

RurangirwaJ

KordariP

2000 Nonpolar inactivation of the hypervariable streptococcal inhibitor of complement gene (sic) in serotype M1 Streptococcus pyogenes significantly decreases mouse mucosal colonization. Infect Immun 68 535 542

69. GryllosI

LevinJC

WesselsMR

2003 The CsrR/CsrS two-component system of group A Streptococcus responds to environmental Mg2+. Proc Natl Acad Sci USA 100 4227 4232

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