Host Cell Invasion and Virulence in Sepsis Is Facilitated by the Multiple Repeats within FnBPA

English version České info

Entry of Staphylococcus aureus into the bloodstream can lead to metastatic abscess formation and infective endocarditis. Crucial to the development of both these conditions is the interaction of S. aureus with endothelial cells. In vivo and in vitro studies have shown that the staphylococcal invasin FnBPA triggers bacterial invasion of endothelial cells via a process that involves fibronectin (Fn) bridging to α₅β₁ integrins. The Fn-binding region of FnBPA usually contains 11 non-identical repeats (FnBRs) with differing affinities for Fn, which facilitate the binding of multiple Fn molecules and may promote integrin clustering. We thus hypothesized that multiple repeats are necessary to trigger the invasion of endothelial cells by S. aureus. To test this we constructed variants of fnbA containing various combinations of FnBRs. In vitro assays revealed that endothelial cell invasion can be facilitated by a single high-affinity, but not low-affinity FnBR. Studies using a nisin-inducible system that controlled surface expression of FnBPA revealed that variants encoding fewer FnBRs required higher levels of surface expression to mediate invasion. High expression levels of FnBPA bearing a single low affinity FnBR bound Fn but did not invade, suggesting that FnBPA affinity for Fn is crucial for triggering internalization. In addition, multiple FnBRs increased the speed of internalization, as did higher expression levels of FnBPA, without altering the uptake mechanism. The relevance of these findings to pathogenesis was demonstrated using a murine sepsis model, which showed that multiple FnBRs were required for virulence. In conclusion, multiple FnBRs within FnBPA facilitate efficient Fn adhesion, trigger rapid bacterial uptake and are required for pathogenesis.

Vyšlo v časopise: Host Cell Invasion and Virulence in Sepsis Is Facilitated by the Multiple Repeats within FnBPA. PLoS Pathog 6(6): e32767. doi:10.1371/journal.ppat.1000964
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1000964

Souhrn

Zdroje

1. LowyFD

1998 Staphylococcus aureus infections. N Engl J Med 339 520 532

2. MenichettiF

2005 Current and emerging serious Gram-positive infections. Clin Microbiol Infect 3 22 28

3. PeacockSJ

de SilvaI

LowyFD

2001 What determines nasal carriage of Staphylococcus aureus? Trends Microbiol 9 605 610

4. FosterTJ

2005 Immune evasion by Staphylococci. Nat Rev Microbiol 3 948 958

5. RooijakkersSH

van KesselKP

van StrijpJA

2005 Staphylococcal innate immune evasion. Trends Microbiol 13 596 601

6. PettiCA

FowlerVGJr

2002 Staphylococcus aureus bacteremia and endocarditis. Infect Dis Clin North Am 16 413 435

7. FowlerVGJr

MiroJM

HoenB

CabellCH

AbrutynE

2005 Staphylococcus aureus endocarditis: a consequence of medical progress. JAMA 293 3012 3021

8. BeynonRP

BahlVK

PrendergastBD

2006 Infective endocarditis. BMJ 333 334 339

9. HillEE

HerijgersP

HerregodsMC

PeetermansWE

2006 Evolving trends in infective endocarditis. Clin Microbiol Infect 12 5 12

10. BashoreTM

CabellC

FowlerVJr

2006 Update on infective endocarditis. Curr Probl Cardiol 31 274 352

11. ChorianopoulosE

BeaF

KatusHA

FreyN

2009 The role of endothelial cell biology in endocarditis. Cell Tissue Res 335 153 163

12. QueYA

HaefligerJA

PirothL

FrançoisP

WidmerE

2005 Fibrinogen and fibronectin-binding cooperate for valve infection and invasion in Staphylococcus aureus experimental endocarditis. J Exp Med 201 1627 1635

13. PirothL

QueYA

WidmerE

PanchaudA

PiuS

2008 The fibrinogen- and fibronectin-binding domains of Staphylococcus aureus fibronectin-binding protein A synergistically promote endothelial invasion and experimental endocarditis. Infect Immun 76 3824 3831

14. PalmqvistN

FosterT

FitzgeraldJR

JosefssonE

TarkowskiA

2005 Fibronectin-binding proteins and fibrinogen-binding clumping factors play distinct roles in staphylococcal arthitis and systemic inflammation. J Infect Dis 191 791 798

15. HeyingR

van de GevelJ

QueYA

MoreillonP

BeekhuizenH

2007 Fibronectin-binding proteins and clumping factor A in Staphylococcus aureus experimental endocarditis: FnBpA is sufficient to activate human endothelial cells. Thomb Haemost 97 617 626

16. HeyingR

van de GevelJ

QueYA

PirothL

MoreillonP

2009 Contribution of (sub)domains of Staphylococcus aureus fibronectin-binding protein to the proinflammatory and procoagulant response of human vascular endothelial cells. Thomb Haemost 101 495 504

17. PeacockSJ

FosterTJ

CameronBJ

BerendtAR

1999 Bacterial fibronectin-binding proteins and endothelial cell surface fibronectin mediate adherence of Staphylococcus aureus to resting human endothelial cells. Microbiology 145 3477 3486

18. SinhaB

FrançoisPP

NüsseO

FotiM

HartfordOM

1999 Fibronectin-binding protein acts as Staphylococcus aureus invasin via fibronectin bridging to integrin α5β1. Cell Microbiol 1 101 117

19. MasseyRC

KantzanouMN

FowlerT

DayNP

SchofieldK

2001 Fibronectin-binding protein A of Staphylococcus aureus has multiple, substituting, binding regions that mediate adherence to Fn and invasion of endothelial cells. Cell Microbiol 3 839 851

20. SinhaB

FrancoisP

QueYA

HussainM

HeilmannC

2000 Heterologously expressed Staphylococcus aureus fibronectin-binding proteins are sufficient for invasion of host cells. Infect Immun 68 6871 6878

21. KerdudouS

LaschkeMW

SinhaB

PreissnerKT

MengerMD

2006 Fibronectin-binding proteins contribute to the adherence of Staphylococcus aureus to intact endothelium in vivo. Thomb Haemost 96 183 189

22. SinhaB

HerrmannM

2005 Mechanism and consequences of invasion of endothelial cells by Staphylococcus aureus. Thomb Haemost 94 266 277

23. IsbergRR

Tran Van NhieuG

1994 Binding and internalization of microorganisms by integrin receptors. Trends Microbiol 2 10 14

24. Nitsche-SchmitzDP

RohdeM

ChhatwalGS

2007 Invasion mechanisms of Gram-positive pathogenic cocci. Thomb Haemost 98 488 496

25. AgererF

LuxS

MichelA

RohdeM

OhlsenK

2005 Cellular invasion by Staphylococcus aureus reveals a functional link between focal adhesion kinase and cortactin in integrin-mediated internalization. J Cell Sci 118 2189 2200

26. SchöderA

SchöderB

RoppenserB

LinderS

Sinha

2006 Staphylococcus aureus fibronectin-binding protein-A induces motile attachment sites and complex actin remodeling in living endothelial cells. Mol Biol Cell 17 5198 5210

27. PeacockSJ

DayNP

ThomasMG

BerendtAR

FosterTJ

2000 Clinical isolates of Staphylococcus aureus exhibit diversity in fnb genes and adhesion to human fibronectin. J Infect 41 23 31

28. WannER

GurusiddappaS

HookMJ

2000 The Fn-binding MSCRAMM FnbpA of Staphylococcus aureus is a bifunctional protein that also binds to fibrinogen. Biol Chem 275 13863 13871

29. RocheFM

DownerR

KeaneF

SpezialeP

ParkPW

2004 The N-terminal A domain of fibronectin-binding proteins A and B promotes adhesion of Staphylococcus aureus to elastin. J Biol Chem 279 38433 38440

30. Schwarz-LinekU

WernerJM

PickfordAR

GurusiddappaS

KimJH

2003 Pathogenic bacteria attach to human fibronectin though a tandem beta-zipper. Nature 423 177 181

31. MeenanNA

VisaiL

ValtulinaV

Schwarz-LinekU

NorrisNC

2007 The tandem beta-zipper model defines high affinity fibronectin-binding repeats within Staphylococcus aureus FnBPA. J Biol Chem 282 25893 25902

32. FrömanG

SwitalskiLM

SpezialeP

HöökMJ

1987 Isolation and characterization of a fibronectin receptor from Staphylococcus aureus. Biol Chem 262 6564 6571

33. BinghamRJ

Rudiño-PiñeraE

MeenanNA

Schwarz-LinekU

TurkenburgJP

2008 Crystal structures of fibronectin-binding sites from Staphylococcus aureus FnBPA in complex with Fn domains. Proc Natl Acad Sci U S A 105 12254 12258

34. GreeneC

McDevittD

FrancoisP

VaudauxPE

LewDP

1995 Adhesion properties of mutants of Staphylococcus aureus defective in fibronectin-binding proteins and studies on the expression of fnb genes. Mol Microbiol 17 1143 1152

35. HirtH

ErlandsenSL

DunnyGM

2000 Heterologous inducible expression of Enterococcus faecalis pCF10 aggregation substance asc10 in Lactococcus lactis and Streptococcus gordonii contributes to cell hydrophobicity and adhesion to fibrin. J Bacteriol 182 2299 2306

36. BryanEM

BaeT

KleerebezemM

DunnyGM

2000 Improved vectors for nisin-controlled expression in gram-positive bacteria. Plasmid 44 183 190

37. LoughmanA

FitzgeraldJR

BrennanMP

HigginsJ

DownerR

2005 Roles for fibrinogen, immunoglobulin and complement in platelet activation promoted by Staphylococcus aureus clumping factor A. Mol Microbiol 57 804 818

38. HylandKA

WangB

ClearyPP

2007 Protein F1 and Streptococcus pyogenes resistance to phagocytosis. Infect Immun 75 3188 3191

39. GarzoniC

KelleyWL

2009 Staphylococcus aureus: new evidence for intracellular persistence. Trends Microbiol 17 59 65

40. SendiP

ProctorRA

2009 Staphylococcus aureus as an intracellular pathogen: the role of small colony variants. Trends Microbiol 17 54 58

41. JosefssonE

HigginsJ

FosterTJ

TarkowskiA

2008 Fibrinogen binding sites P336 and Y338 of clumping factor A are crucial for Staphylococcus aureus virulence. PLoS ONE 3 e2206

42. EdgellCJ

McDonaldCC

GrahamJB

1983 Permanent cell line expressing human factor VIII-related antigen established by hybridization. Proc Natl Acad Sci U S A 80 3734 3737

43. SambrookJ

FritschEF

ManiatisT

1989 Molecular Cloning: a Laboratory Manual, 2nd edn. Cold Spring Harbor,, NY Cold Spring Harbor Laboratory

44. JakubovicsNS

StrömbergN

van DolleweerdCJ

KellyCG

JenkinsonHF

2005 Differential binding specificities of oral streptococcal antigen I/II family adhesins for human or bacterial ligands. Mol Microbiol 55 1591 1605

45. KreiswirthBN

LöfdahlS

BetleyMJ

O'ReillyM

SchlievertPM

1983 The toxic shock syndrome exotoxin structural gene is not detectably transmitted by a prophage. Nature 305 709 712

46. NovickR

1967 Properties of a cryptic high-frequency transducing phage in Staphylococcus aureus. Virology 33 155 166

47. HartleibJ

KöhlerN

DickinsonRB

ChhatwalGS

SixmaJJ

2000 Protein A is the von Willebrand factor binding protein on Staphylococcus aureus. Blood 96 2149 2156

48. BremellT

LangeS

YacoubA

RydénC

TarkowskiA

1991 Experimental Staphylococcus aureus arthitis in mice. Infect Immun 59 2615 2623

49. HartfordO

O'BrienL

SchofieldK

WellsJ

FosterTJ

2001 The Fbe (SdrG) protein of Staphylococcus epidermidis HB promotes bacterial adherence to fibrinogen. Microbiology 147 2545 2552

50. KuipersOP

BeerthuyzenMM

SiezenRJ

De VosWM

1993 Characterization of the nisin gene cluster nisABTCIPR of Lactococcus lactis. Requirement of expression of the nisA and nisI genes for development of immunity. Eur J Biochem 216 281 291