#PAGE_PARAMS# #ADS_HEAD_SCRIPTS# #MICRODATA#

Type III Secretion Protein MxiI Is Recognized by Naip2 to Induce Nlrc4 Inflammasome Activation Independently of Pkcδ


Recognition of intracellular pathogenic bacteria by members of the nucleotide-binding domain and leucine-rich repeat containing (NLR) family triggers immune responses against bacterial infection. A major response induced by several Gram-negative bacteria is the activation of caspase-1 via the Nlrc4 inflammasome. Upon activation, caspase-1 regulates the processing of proIL-1β and proIL-18 leading to the release of mature IL-1β and IL-18, and induction of pyroptosis. The activation of the Nlrc4 inflammasome requires the presence of an intact type III or IV secretion system that mediates the translocation of small amounts of flagellin or PrgJ-like rod proteins into the host cytosol to induce Nlrc4 activation. Using the Salmonella system, it was shown that Naip2 and Naip5 link flagellin and the rod protein PrgJ, respectively, to Nlrc4. Furthermore, phosphorylation of Nlrc4 at Ser533 by Pkcδ was found to be critical for the activation of the Nlrc4 inflammasome. Here, we show that Naip2 recognizes the Shigella T3SS inner rod protein MxiI and induces Nlrc4 inflammasome activation. The expression of MxiI in primary macrophages was sufficient to induce pyroptosis and IL-1β release, which were prevented in macrophages deficient in Nlrc4. In the presence of MxiI or Shigella infection, MxiI associated with Naip2, and Naip2 interacted with Nlrc4. siRNA-mediated knockdown of Naip2, but not Naip5, inhibited Shigella-induced caspase-1 activation, IL-1β maturation and Asc pyroptosome formation. Notably, the Pkcδ kinase was dispensable for caspase-1 activation and secretion of IL-1β induced by Shigella or Salmonella infection. These results indicate that activation of caspase-1 by Shigella is triggered by the rod protein MxiI that interacts with Naip2 to induce activation of the Nlrc4 inflammasome independently of the Pkcδ kinase.


Vyšlo v časopise: Type III Secretion Protein MxiI Is Recognized by Naip2 to Induce Nlrc4 Inflammasome Activation Independently of Pkcδ. PLoS Pathog 10(2): e32767. doi:10.1371/journal.ppat.1003926
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1003926

Souhrn

Recognition of intracellular pathogenic bacteria by members of the nucleotide-binding domain and leucine-rich repeat containing (NLR) family triggers immune responses against bacterial infection. A major response induced by several Gram-negative bacteria is the activation of caspase-1 via the Nlrc4 inflammasome. Upon activation, caspase-1 regulates the processing of proIL-1β and proIL-18 leading to the release of mature IL-1β and IL-18, and induction of pyroptosis. The activation of the Nlrc4 inflammasome requires the presence of an intact type III or IV secretion system that mediates the translocation of small amounts of flagellin or PrgJ-like rod proteins into the host cytosol to induce Nlrc4 activation. Using the Salmonella system, it was shown that Naip2 and Naip5 link flagellin and the rod protein PrgJ, respectively, to Nlrc4. Furthermore, phosphorylation of Nlrc4 at Ser533 by Pkcδ was found to be critical for the activation of the Nlrc4 inflammasome. Here, we show that Naip2 recognizes the Shigella T3SS inner rod protein MxiI and induces Nlrc4 inflammasome activation. The expression of MxiI in primary macrophages was sufficient to induce pyroptosis and IL-1β release, which were prevented in macrophages deficient in Nlrc4. In the presence of MxiI or Shigella infection, MxiI associated with Naip2, and Naip2 interacted with Nlrc4. siRNA-mediated knockdown of Naip2, but not Naip5, inhibited Shigella-induced caspase-1 activation, IL-1β maturation and Asc pyroptosome formation. Notably, the Pkcδ kinase was dispensable for caspase-1 activation and secretion of IL-1β induced by Shigella or Salmonella infection. These results indicate that activation of caspase-1 by Shigella is triggered by the rod protein MxiI that interacts with Naip2 to induce activation of the Nlrc4 inflammasome independently of the Pkcδ kinase.


Zdroje

1. FranchiL, Muñoz-PlanilloR, NúñezG (2012) Sensing and reacting to microbes through the inflammasomes. Nat immunol 13: 325–332.

2. RathinamVA, VanajaSK, FitzgeraldKA (2012) Regulation of inflammasome signaling. Nat Immunol 13: 333–2.

3. BrozP, MonackDM (2011) Molecular mechanisms of inflammasome activation during microbial infections. Immunol Rev 243: 174–90.

4. FranchiL, AmerA, Body-MalapelM, KannegantiTD, OzörenN, et al. (2006) Cytosolic flagellin requires Ipaf for activation of caspase-1 and interleukin 1beta in Salmonella-infected macrophages. Nat Immunol 7: 576–582.

5. FranchiL, EigenbrodT, Muñoz-PlanilloR, NuñezG (2009) The inflammasome: a caspase-1-activation platform that regulates immune responses and disease pathogenesis. Nat immunol 10: 241–247.

6. LamkanfiM, DixitVM (2012) Inflammasomes and their roles in health and disease. Annu Rev Cell Dev Biol 28: 137–61.

7. AshidaH, OgawaM, KimM, MimuroH, SasakawaC (2011) Bacteria and host interactions in the gut epithelial barrier. Nat Chem Biol 8: 36–45.

8. AshidaH, OgawaM, KimM, SuzukiS, SanadaT, et al. (2011) Shigella deploy multiple countermeasures against host innate immune responses. Curr Opin Microbiol 14: 16–23.

9. MiaoEA, MaoDP, YudkovskyN, BonneauR, LorangCG, et al. (2010) Innate immune detection of the type III secretion apparatus through the NLRC4 inflammasome. Proc Natl Acad Sci USA 107: 3076–3080.

10. KofoedEM, VanceRE (2011) Innate immune recognition of bacterial ligands by NAIPs determines inflammasome specificity. Nature 477: 592–595.

11. ZhaoY, YangJ, ShiJ, GongY, LuQ, et al. (2011) The NLRC4 inflammasome receptors for bacterial flagellin and type III secretion apparatus. Nature 477: 596–600.

12. HalffEF, DiebolderCA, VersteegM, SchoutenA, BrondijkTH, et al. (2012) Formation and structure of a NAIP5-NLRC4 inflammasome induced by direct interactions with conserved N- and C-terminal regions of flagellin. J Biol Chem 287: 38460–72.

13. QuY, MisaghiS, Izrael-TomasevicA, NewtonK, GilmourLL, et al. (2012) Phosphorylation of NLRC4 is critical for inflammasome activation. Nature 490: 539–42.

14. SchroederGN, HilbiH (2008) Molecular pathogenesis of Shigella spp.: controlling host cell signaling, invasion, and death by type III secretion. Clin Microbiol Rev 21: 134–56.

15. OgawaM, HandaY, AshidaH, SuzukiM, SasakawaC (2008) The versatility of Shigella effectors. Nat Rev Microbiol 6: 11–6.

16. MarlovitsTC, KuboriT, SukhanA, ThomasDR, GalánJE, et al. (2004) Structural insights into the assembly of the type III secretion needle complex. Science 306: 1040–1042.

17. SaniM, AllaouiA, FusettiF, OostergetelGT, KeegstraW, et al. (2007) Structural organization of the needle complex of the type III secretion apparatus of Shigella flexneri. Micron 38: 291–301.

18. SuzukiT, FranchiL, TomaC, AshidaH, OgawaM, et al. (2007) Differential regulation of caspase-1 activation, pyroptosis, and autophagy via Ipaf and ASC in Shigella infected macrophages. PLoS Pathog 3: 1082–1091.

19. MagdalenaJ, HachaniA, ChamekhM, JouihriN, GounonP, et al. (2002) Spa32 Regulates a Switch in Substrate Specificity of the Type III Secreton of Shigella flexneri from Needle Components to Ipa proteins. J Bacteriol 184: 3433–3441.

20. JouihriN, SoryMP, PageAL, GounonP, ParsotC, et al. (2003) MxiK and MxiN interact with the Spa47 ATPase and are required for transit of the needle components MxiH and MxiI, but not of Ipa proteins, through the type III secretion apparatus of Shigella flexneri. Mol Microbiol 49: 755–767.

21. BotteauxA, KayathCA, PageAL, JouihriN, SaniM, et al. (2010) The 33 carboxyl-terminal residues of Spa40 orchestrate the multi-step assembly process of the type III secretion needle complex in Shigella flexneri. Microbiol 156: 2807–2817.

22. KimbroughTG, MillerSI (2000) Contribution of Salmonella typhimurium type III secretion components to needle complex formation. Proc Natl Acad Sci USA 97: 11008–11013.

23. MiaoEA, RajanJV (2011) Salmonella and Caspase-1: A complex Interplay of Detection and Evasion. Front Microbiol 2: 85.

24. WillinghamSB, BergstralhDT, O'ConnorW, MorrisonAC, TaxmanDJ, et al. (2007) Microbial pathogen-induced necrotic cell death mediated by the inflammasome components CIAS1/Cryopyrin/NLRP3 and ASC. Cell Host Microbe 2: 147–159.

25. Fernandes-AlnemriT, WuJ, YuJW, DattaP, MillerB, eyal (2007) The pyroptosome: a supramolecular assembly of ASC dimers mediating inflammatory cell death via caspase-1 activation. Cell Death Differ 14: 1590–604.

26. Fernandes-AlnemriT, AlnemriES (2008) Assembly, purification, and assay of the activity of the ASC pyroptosome. Methods Enzymol 442: 251–270.

27. JulianaC, Fernandes-AlnemriT, WuJ, DattaP, SolorzanoL, et al. (2010) Anti-inflammatory compounds parthenolide and bay 11-7082 are direct inhibitors of the inflammasome. J Biol Chem 285: 9792–802.

28. BrownGE, StewartMQ, LiuH, HaVL, YaffeMB (2003) A novel assay system implicates PtdIns(3,4)P(2), PtdIns(3)P, and PKC delta in intracellular production of reactive oxygen species by the NADPH oxidase. Mol Cell 11: 35–47.

29. MeissnerF, SegerRA, MoshousD, FischerA, ReichenbachJ, et al. (2010) Inflammasome activation in NADPH oxidase defective mononuclear phagocytes from patients with chronic granulomatous disease. Blood 116: 1570–1573.

30. BasuA (2003) Involvement of protein kinase C-delta in DNA damage-induced apoptosis. J Cell Mol Med 7: 341–50.

31. BrodieC, BlumbergPM (2003) Regulation of cell apoptosis by protein kinase c delta. Apoptosis 8: 19–27.

32. ReylandME, AndersonSM, MatassaAA, BarzenKA, QuissellDO (1999) Protein kinase C delta is essential for etoposide-induced apoptosis in salivary gland acinar cells. J Biol Chem 274: 19115–23.

33. MariathasanS, HewtonK, MonackDM, VucicD, FrenchDM, et al. (2004) Differential activation of the inflammasome by caspase-1 adaptors ASC and Ipaf. Nature 430: 213–218.

34. KannegantiTD, Body-MalapelM, AmerA, ParkJH, WhitfieldJ, et al. (2006) Critical role for cryopyrin/Nalp3 in activation of caspase-1 in response to viral infection and double-stranded RNA. J Biol Chem 281: 36560–36568.

35. OzorenN, MasumotoJ, FranchiL, KannegantiTD, Body-MalapelM, et al. (2006) Distinct roles of TLR2 and the adaptor ASC in IL-1beta/IL-18 secretion in response to Listeria monocytogenes. J Immunol 176: 4337–4342.

36. SasakawaC, KamataK, SakaiT, MurayamaY, MakinoS, et al. (1986) Molecular alteration of the 140-megadalton plasmid associated with loss of virulence and congo red binding activity in Shigella flexneri. Infect Immun 51: 470–475.

37. WataraiM, TobeT, YoshikawaM, SasakawaC (1995) Contact of Shigella with host cells triggers release of Ipa invasins and is an essential function of invasiveness. EMBO J 14: 2461–2470.

38. KodamaC, MatsuiH (2004) Salmonella flagellin is not a dominant protective antigen in oral immunization with attenuated live vaccine strains. Infect Immun 72: 2449–2451.

39. FranchiL, KamadaN, NakamuraY, BurberryA, KuffaP, et al. (2012) NLRC4-driven production of IL-1β discriminates between pathogenic and commensal bacteria and promotes host intestinal defense. Nat Immunol 13: 449–456.

40. SuzukiY, Yoshitomo-NakagawaK, MaruyamaK, SuyamaA, SuganoS (1997) Construction and characterization of a full length-enriched and a 5′-end-enriched cDNA library. Gene 200: 149–156.

41. AdamiC, BrundaMJ, PalleroniAV (1993) In vivo immortalization of murine peritoneal macrophages: a new rapid and efficient method for obtaining macrophage cell lines. J Leukoc Biol 53: 475–8.

Štítky
Hygiena a epidemiológia Infekčné lekárstvo Laboratórium

Článok vyšiel v časopise

PLOS Pathogens


2014 Číslo 2
Najčítanejšie tento týždeň
Najčítanejšie v tomto čísle
Kurzy

Zvýšte si kvalifikáciu online z pohodlia domova

Získaná hemofilie - Povědomí o nemoci a její diagnostika
nový kurz

Eozinofilní granulomatóza s polyangiitidou
Autori: doc. MUDr. Martina Doubková, Ph.D.

Všetky kurzy
Prihlásenie
Zabudnuté heslo

Zadajte e-mailovú adresu, s ktorou ste vytvárali účet. Budú Vám na ňu zasielané informácie k nastaveniu nového hesla.

Prihlásenie

Nemáte účet?  Registrujte sa

#ADS_BOTTOM_SCRIPTS#