NOD2, the nucleotide-binding domain and leucine-rich repeat containing gene family (NLR) member 2 is involved in mediating antimicrobial responses. Dysfunctional NOD2 activity can lead to severe inflammatory disorders, but the regulation of NOD2 is still poorly understood. Recently, proteins of the tripartite motif (TRIM) protein family have emerged as regulators of innate immune responses by acting as E3 ubiquitin ligases. We identified TRIM27 as a new specific binding partner for NOD2. We show that NOD2 physically interacts with TRIM27 via the nucleotide-binding domain, and that NOD2 activation enhances this interaction. Dependent on functional TRIM27, ectopically expressed NOD2 is ubiquitinated with K48-linked ubiquitin chains followed by proteasomal degradation. Accordingly, TRIM27 affects NOD2-mediated pro-inflammatory responses. NOD2 mutations are linked to susceptibility to Crohn's disease. We found that TRIM27 expression is increased in Crohn's disease patients, underscoring a physiological role of TRIM27 in regulating NOD2 signaling. In HeLa cells, TRIM27 is partially localized in the nucleus. We revealed that ectopically expressed NOD2 can shuttle to the nucleus in a Walker A dependent manner, suggesting that NOD2 and TRIM27 might functionally cooperate in the nucleus. We conclude that TRIM27 negatively regulates NOD2-mediated signaling by degradation of NOD2 and suggest that TRIM27 could be a new target for therapeutic intervention in NOD2-associated diseases.
References
[1]
Ting JP, Duncan JA, Lei Y (2010) How the noninflammasome NLRs function in the innate immune system. Science 327: 286–290.
[2]
Chen G, Shaw MH, Kim YG, Nunez G (2009) NOD-like receptors: role in innate immunity and inflammatory disease. Annu Rev Pathol 4: 365–398.
[3]
Magalhaes JG, Sorbara MT, Girardin SE, Philpott DJ (2011) What is new with Nods? Curr Opin Immunol 23: 29–34.
[4]
Kufer TA, Kremmer E, Banks DJ, Philpott DJ (2006) Role for erbin in bacterial activation of Nod2. Infect Immun 74: 3115–3124.
[5]
Barnich N, Aguirre JE, Reinecker HC, Xavier R, Podolsky DK (2005) Membrane recruitment of NOD2 in intestinal epithelial cells is essential for nuclear factor-{kappa}B activation in muramyl dipeptide recognition. J Cell Biol 170: 21–26.
[6]
Legrand-Poels S, Kustermans G, Bex F, Kremmer E, Kufer TA, et al. (2007) Modulation of Nod2-dependent NF-kappaB signaling by the actin cytoskeleton. J Cell Sci 120: 1299–1310.
[7]
Philpott DJ, Girardin SE (2010) Nod-like receptors: sentinels at host membranes. Curr Opin Immunol 22: 428–434.
[8]
Ogura Y, Inohara N, Benito A, Chen FF, Yamaoka S, et al. (2001) Nod2, a Nod1/Apaf-1 family member that is restricted to monocytes and activates NF-kappaB. J Biol Chem 276: 4812–4818.
[9]
Kobayashi K, Inohara N, Hernandez LD, Galan JE, Nunez G, et al. (2002) RICK/Rip2/CARDIAK mediates signalling for receptors of the innate and adaptive immune systems. Nature 416: 194–199.
[10]
Kobayashi KS, Chamaillard M, Ogura Y, Henegariu O, Inohara N, et al. (2005) Nod2-dependent regulation of innate and adaptive immunity in the intestinal tract. Science 307: 731–734.
[11]
Borzutzky A, Fried A, Chou J, Bonilla FA, Kim S, et al. (2009) NOD2-associated diseases: Bridging innate immunity and autoinflammation. Clin Immunol.
[12]
McDonald C, Chen FF, Ollendorff V, Ogura Y, Marchetto S, et al. (2005) A role for Erbin in the regulation of Nod2-dependent NF-kappaB signaling. J Biol Chem 280: 40301–40309.
Bielig H, Zurek B, Kutsch A, Menning M, Philpott DJ, et al. (2009) A function for AAMP in Nod2-mediated NF-kappaB activation. Mol Immunol 46: 2647–54.
[15]
Sardiello M, Cairo S, Fontanella B, Ballabio A, Meroni G (2008) Genomic analysis of the TRIM family reveals two groups of genes with distinct evolutionary properties. BMC Evol Biol 8: 225.
[16]
Rajsbaum R, Stoye JP, O'Garra A (2008) Type I interferon-dependent and -independent expression of tripartite motif proteins in immune cells. Eur J Immunol 38: 619–630.
[17]
Carthagena L, Bergamaschi A, Luna JM, David A, Uchil PD, et al. (2009) Human TRIM gene expression in response to interferons. PLoS One 4: e4894.
[18]
Ozato K, Shin DM, Chang TH, Morse HC III (2008) TRIM family proteins and their emerging roles in innate immunity. Nat Rev Immunol 8: 849–860.
[19]
McNab FW, Rajsbaum R, Stoye JP, O'Garra A (2011) Tripartite-motif proteins and innate immune regulation. Curr Opin Immunol 23: 46–56.
[20]
Bailly V, Lauder S, Prakash S, Prakash L (1997) Yeast DNA repair proteins Rad6 and Rad18 form a heterodimer that has ubiquitin conjugating, DNA binding, and ATP hydrolytic activities. J Biol Chem 272: 23360–23365.
[21]
Deshaies RJ, Joazeiro CA (2009) RING domain E3 ubiquitin ligases. Annu Rev Biochem 78: 399–434.
[22]
French FMF Consortium (1997) A candidate gene for familial Mediterranean fever. Nat Genet 17: 25–31.
[23]
International FMF Consortium (1997) Ancient missense mutations in a new member of the RoRet gene family are likely to cause familial Mediterranean fever. The International FMF Consortium. Cell 90: 797–807.
[24]
Harley JB, Alexander EL, Bias WB, Fox OF, Provost TT, et al. (1986) Anti-Ro (SS-A) and anti-La (SS-B) in patients with Sjogren's syndrome. Arthritis Rheum 29: 196–206.
[25]
Ishii T, Ohnuma K, Murakami A, Takasawa N, Yamochi T, et al. (2003) SS-A/Ro52, an autoantigen involved in CD28-mediated IL-2 production. J Immunol 170: 3653–3661.
[26]
Nakayama EE, Shioda T (2010) Anti-retroviral activity of TRIM5 alpha. Rev Med Virol 20: 77–92.
[27]
Hasegawa N, Iwashita T, Asai N, Murakami H, Iwata Y, et al. (1996) A RING finger motif regulates transforming activity of the rfp/ret fusion gene. Biochem Biophys Res Commun 225: 627–631.
[28]
Napolitano LM, Jaffray EG, Hay RT, Meroni G (2011) Functional interactions between ubiquitin E2 enzymes and TRIM proteins. Biochem J 434: 309–319.
[29]
Chu Y, Yang X (2011) SUMO E3 ligase activity of TRIM proteins. Oncogene 30: 1108–1116.
[30]
Tezel G, Nagasaka T, Iwahashi N, Asai N, Iwashita T, et al. (1999) Different nuclear/cytoplasmic distributions of RET finger protein in different cell types. Pathol Int 49: 881–886.
[31]
Cao T, Borden KL, Freemont PS, Etkin LD (1997) Involvement of the rfp tripartite motif in protein-protein interactions and subcellular distribution. J Cell Sci 110(Pt 14): 1563–1571.
[32]
Cao T, Duprez E, Borden KL, Freemont PS, Etkin LD (1998) Ret finger protein is a normal component of PML nuclear bodies and interacts directly with PML. J Cell Sci 111(Pt 10): 1319–1329.
[33]
Matsuura T, Shimono Y, Kawai K, Murakami H, Urano T, et al. (2005) PIAS proteins are involved in the SUMO-1 modification, intracellular translocation and transcriptional repressive activity of RET finger protein. Exp Cell Res 308: 65–77.
[34]
Shimono Y, Murakami H, Hasegawa Y, Takahashi M (2000) RET finger protein is a transcriptional repressor and interacts with enhancer of polycomb that has dual transcriptional functions. J Biol Chem 275: 39411–39419.
[35]
Bloor AJ, Kotsopoulou E, Hayward P, Champion BR, Green AR (2005) RFP represses transcriptional activation by bHLH transcription factors. Oncogene 24: 6729–6736.
[36]
Zha J, Han KJ, Xu LG, He W, Zhou Q, et al. (2006) The Ret finger protein inhibits signaling mediated by the noncanonical and canonical IkappaB kinase family members. J Immunol 176: 1072–1080.
[37]
Dho SH, Kwon KS (2003) The Ret finger protein induces apoptosis via its RING finger-B box-coiled-coil motif. J Biol Chem 278: 31902–31908.
[38]
Patel CA, Ghiselli G (2005) The RET finger protein interacts with the hinge region of SMC3. Biochem Biophys Res Commun 330: 333–340.
[39]
Reymond A, Meroni G, Fantozzi A, Merla G, Cairo S, et al. (2001) The tripartite motif family identifies cell compartments. Embo J 20: 2140–2151.
[40]
Hershko A, Ciechanover A (1998) The ubiquitin system. Annu Rev Biochem 67: 425–479.
[41]
Benko S, Magalhaes JG, Philpott DJ, Girardin SE (2010) NLRC5 limits the activation of inflammatory pathways. J Immunol 185: 1681–1691.
[42]
Cressman DE, Chin KC, Taxman DJ, Ting JP (1999) A defect in the nuclear translocation of CIITA causes a form of type II bare lymphocyte syndrome. Immunity 10: 163–171.
[43]
Harton JA, Cressman DE, Chin KC, Der CJ, Ting JP (1999) GTP binding by class II transactivator: role in nuclear import. Science 285: 1402–1405.
[44]
Meissner TB, Li A, Biswas A, Lee KH, Liu YJ, et al. (2010) NLR family member NLRC5 is a transcriptional regulator of MHC class I genes. Proc Natl Acad Sci U S A 107: 13794–13799.
[45]
Neerincx A, Rodriguez GM, Steimle V, Kufer TA (2012) NLRC5 Controls Basal MHC Class I Gene Expression in an MHC Enhanceosome-Dependent Manner. J Immunol 188: 4940–4950.
[46]
Uehara A, Yang S, Fujimoto Y, Fukase K, Kusumoto S, et al. (2005) Muramyldipeptide and diaminopimelic acid-containing desmuramylpeptides in combination with chemically synthesized Toll-like receptor agonists synergistically induced production of interleukin-8 in a NOD2- and NOD1-dependent manner, respectively, in human monocytic cells in culture. Cell Microbiol 7: 53–61.
[47]
Rhodes DA, de Bono B, Trowsdale J (2005) Relationship between SPRY and B30.2 protein domains. Evolution of a component of immune defence? Immunology 116: 411–417.
[48]
Henry J, Ribouchon M, Depetris D, Mattei M, Offer C, et al. (1997) Cloning, structural analysis, and mapping of the B30 and B7 multigenic families to the major histocompatibility complex (MHC) and other chromosomal regions. Immunogenetics 46: 383–395.
[49]
Kong HJ, Anderson DE, Lee CH, Jang MK, Tamura T, et al. (2007) Cutting edge: autoantigen Ro52 is an interferon inducible E3 ligase that ubiquitinates IRF-8 and enhances cytokine expression in macrophages. J Immunol 179: 26–30.
[50]
Higgs R, Ni Gabhann J, Ben Larbi N, Breen EP, Fitzgerald KA, et al. (2008) The E3 ubiquitin ligase Ro52 negatively regulates IFN-beta production post-pathogen recognition by polyubiquitin-mediated degradation of IRF3. J Immunol 181: 1780–1786.
[51]
Yang K, Shi HX, Liu XY, Shan YF, Wei B, et al. (2009) TRIM21 is essential to sustain IFN regulatory factor 3 activation during antiviral response. J Immunol 182: 3782–3792.
[52]
Gack MU, Shin YC, Joo CH, Urano T, Liang C, et al. (2007) TRIM25 RING-finger E3 ubiquitin ligase is essential for RIG-I-mediated antiviral activity. Nature 446: 916–920.
[53]
Richards N, Schaner P, Diaz A, Stuckey J, Shelden E, et al. (2001) Interaction between pyrin and the apoptotic speck protein (ASC) modulates ASC-induced apoptosis. J Biol Chem 276: 39320–39329.
[54]
Chae JJ, Wood G, Masters SL, Richard K, Park G, et al. (2006) The B30.2 domain of pyrin, the familial Mediterranean fever protein, interacts directly with caspase-1 to modulate IL-1beta production. Proc Natl Acad Sci U S A 103: 9982–9987.
[55]
Chae JJ, Komarow HD, Cheng J, Wood G, Raben N, et al. (2003) Targeted disruption of pyrin, the FMF protein, causes heightened sensitivity to endotoxin and a defect in macrophage apoptosis. Mol Cell 11: 591–604.
[56]
Chae JJ, Cho YH, Lee GS, Cheng J, Liu PP, et al. (2011) Gain-of-Function Pyrin Mutations Induce NLRP3 Protein-Independent Interleukin-1beta Activation and Severe Autoinflammation in Mice. Immunity 34: 755–768.
[57]
Laing KJ, Purcell MK, Winton JR, Hansen JD (2008) A genomic view of the NOD-like receptor family in teleost fish: identification of a novel NLR subfamily in zebrafish. BMC Evol Biol 8: 42.
[58]
van der Aa LM, Levraud JP, Yahmi M, Lauret E, Briolat V, et al. (2009) A large new subset of TRIM genes highly diversified by duplication and positive selection in teleost fish. BMC Biol 7: 7.
[59]
Toniato E, Chen XP, Losman J, Flati V, Donahue L, et al. (2002) TRIM8/GERP RING finger protein interacts with SOCS-1. J Biol Chem 277: 37315–37322.
[60]
Arimoto K, Funami K, Saeki Y, Tanaka K, Okawa K, et al. (2010) Polyubiquitin conjugation to NEMO by triparite motif protein 23 (TRIM23) is critical in antiviral defense. Proc Natl Acad Sci U S A 107: 15856–15861.
[61]
Tsuchida T, Zou J, Saitoh T, Kumar H, Abe T, et al. (2010) The ubiquitin ligase TRIM56 regulates innate immune responses to intracellular double-stranded DNA. Immunity 33: 765–776.
[62]
Hu Y, Mao K, Zeng Y, Chen S, Tao Z, et al. (2010) Tripartite-motif protein 30 negatively regulates NLRP3 inflammasome activation by modulating reactive oxygen species production. J Immunol 185: 7699–7705.
[63]
Shi M, Deng W, Bi E, Mao K, Ji Y, et al. (2008) TRIM30 alpha negatively regulates TLR-mediated NF-kappa B activation by targeting TAB2 and TAB3 for degradation. Nat Immunol 9: 369–377.
[64]
von Mikecz A (2006) The nuclear ubiquitin-proteasome system. J Cell Sci 119: 1977–1984.
Lu D, Lin W, Gao X, Wu S, Cheng C, et al. (2011) Direct ubiquitination of pattern recognition receptor FLS2 attenuates plant innate immunity. Science 332: 1439–1442.
[67]
Steimle V, Otten LA, Zufferey M, Mach B (1993) Complementation cloning of an MHC class II transactivator mutated in hereditary MHC class II deficiency (or bare lymphocyte syndrome). Cell 75: 135–146.
[68]
Zika E, Ting JP (2005) Epigenetic control of MHC-II: interplay between CIITA and histone-modifying enzymes. Curr Opin Immunol 17: 58–64.
[69]
Staehli F, Ludigs K, Heinz LX, Seguin-Estevez Q, Ferrero I, et al. (2012) NLRC5 deficiency selectively impairs MHC class I- dependent lymphocyte killing by cytotoxic T cells. J Immunol 188: 3820–3828.
[70]
Tameling WI, Baulcombe DC (2007) Physical association of the NB-LRR resistance protein Rx with a Ran GTPase-activating protein is required for extreme resistance to Potato virus X. Plant Cell 19: 1682–1694.
[71]
Wirthmueller L, Zhang Y, Jones JD, Parker JE (2007) Nuclear accumulation of the Arabidopsis immune receptor RPS4 is necessary for triggering EDS1-dependent defense. Curr Biol 17: 2023–2029.
[72]
Burch-Smith TM, Schiff M, Caplan JL, Tsao J, Czymmek K, et al. (2007) A novel role for the TIR domain in association with pathogen-derived elicitors. PLoS Biol 5: e68.
[73]
Shen QH, Saijo Y, Mauch S, Biskup C, Bieri S, et al. (2007) Nuclear activity of MLA immune receptors links isolate-specific and basal disease-resistance responses. Science 315: 1098–1103.
[74]
Liu J, Coaker G (2008) Nuclear trafficking during plant innate immunity. Mol Plant 1: 411–422.
[75]
Liu Y, Jin H, Yang KY, Kim CY, Baker B, et al. (2003) Interaction between two mitogen-activated protein kinases during tobacco defense signaling. Plant J 34: 149–160.
[76]
Jin H, Liu Y, Yang KY, Kim CY, Baker B, et al. (2003) Function of a mitogen-activated protein kinase pathway in N gene-mediated resistance in tobacco. Plant J 33: 719–731.
[77]
Zurek B, Proell M, Wagner RN, Schwarzenbacher R, Kufer TA (2012) Mutational analysis of human NOD1 and NOD2 NACHT domains reveals different modes of activation. Innate Immun 18: 100–11.
[78]
Lecat A, Piette J, Legrand-Poels S (2010) The protein Nod2: an innate receptor more complex than previously assumed. Biochem Pharmacol 80: 2021–2031.
[79]
Kufer TA (2008) Signal transduction pathways used by NLR-type innate immune receptors. Mol Biosyst 4: 380–386.
[80]
Sabbah A, Chang TH, Harnack R, Frohlich V, Tominaga K, et al. (2009) Activation of innate immune antiviral responses by Nod2. Nat Immunol 10: 1073–1080.
[81]
Oh HM, Lee HJ, Seo GS, Choi EY, Kweon SH, et al. (2005) Induction and localization of NOD2 protein in human endothelial cells. Cell Immunol 237: 37–44.
[82]
Rosenstiel P, Fantini M, Brautigam K, Kuhbacher T, Waetzig GH, et al. (2003) TNF-alpha and IFN-gamma regulate the expression of the NOD2 (CARD15) gene in human intestinal epithelial cells. Gastroenterology 124: 1001–1009.
[83]
Gutierrez O, Pipaon C, Inohara N, Fontalba A, Ogura Y, et al. (2002) Induction of Nod2 in myelomonocytic and intestinal epithelial cells via nuclear factor-kappa B activation. J Biol Chem 277: 41701–41705.
[84]
Ogura Y, Lala S, Xin W, Smith E, Dowds TA, et al. (2003) Expression of NOD2 in Paneth cells: a possible link to Crohn's ileitis. Gut 52: 1591–1597.
[85]
Kufer TA, Kremmer E, Adam AC, Philpott DJ, Sansonetti PJ (2008) The pattern-recognition molecule Nod1 is localized at the plasma membrane at sites of bacterial interaction. Cell Microbiol 10: 477–486.
[86]
Lim KL, Chew KC, Tan JM, Wang C, Chung KK, et al. (2005) Parkin mediates nonclassical, proteasomal-independent ubiquitination of synphilin-1: implications for Lewy body formation. J Neurosci 25: 2002–2009.
[87]
Gyuris J, Golemis E, Chertkov H, Brent R (1993) Cdi1, a human G1 and S phase protein phosphatase that associates with Cdk2. Cell 75: 791–803.
[88]
Rasband WS (2007) ImageJ. Bethesda, Maryland, USA: U.S. National Institutes of Health.
