Abstract
Innate immunity is a fundamental defence response that depends on evolutionarily conserved pattern recognition receptors for sensing infections or danger signals1,2. Nucleotide-binding and oligomerization domain (NOD) proteins are cytosolic pattern-recognition receptors of paramount importance in the intestine, and their dysregulation is associated with inflammatory bowel disease3,4. They sense peptidoglycans from commensal microorganisms and pathogens and coordinate signalling events that culminate in the induction of inflammation and anti-microbial responses2. However, the signalling mechanisms involved in this process are not fully understood. Here, using genome-wide RNA interference, we identify candidate genes that modulate the NOD1 inflammatory response in intestinal epithelial cells. Our results reveal a significant crosstalk between innate immunity and apoptosis and identify BID, a BCL2 family protein, as a critical component of the inflammatory response. Colonocytes depleted of BID or macrophages from Bid−/− mice are markedly defective in cytokine production in response to NOD activation. Furthermore, Bid−/− mice are unresponsive to local or systemic exposure to NOD agonists or their protective effect in experimental colitis. Mechanistically, BID interacts with NOD1, NOD2 and the IκB kinase (IKK) complex, impacting NF-κB and extracellular signal-regulated kinase (ERK) signalling. Our results define a novel role of BID in inflammation and immunity independent of its apoptotic function, furthering the mounting evidence of evolutionary conservation between the mechanisms of apoptosis and immunity.
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References
Garrett, W. S., Gordon, J. I. & Glimcher, L. H. Homeostasis and inflammation in the intestine. Cell 140, 859–870 (2010)
Takeuchi, O. & Akira, S. Pattern recognition receptors and inflammation. Cell 140, 805–820 (2010)
Cho, J. H. The genetics and immunopathogenesis of inflammatory bowel disease. Nature Rev. Immunol. 8, 458–466 (2008)
Turner, J. R. Intestinal mucosal barrier function in health and disease. Nature Rev. Immunol. 9, 799–809 (2009)
Girardin, S. E. et al. Nod1 detects a unique muropeptide from Gram-negative bacterial peptidoglycan. Science 300, 1584–1587 (2003)
Kim, J. G., Lee, S. J. & Kagnoff, M. F. Nod1 is an essential signal transducer in intestinal epithelial cells infected with bacteria that avoid recognition by toll-like receptors. Infect. Immun. 72, 1487–1495 (2004)
Masumoto, J. et al. Nod1 acts as an intracellular receptor to stimulate chemokine production and neutrophil recruitment in vivo . J. Exp. Med. 203, 203–213 (2006)
Chen, C. M., Gong, Y., Zhang, M. & Chen, J. J. Reciprocal cross-talk between Nod2 and TAK1 signaling pathways. J. Biol. Chem. 279, 25876–25882 (2004)
Kobayashi, K. S. et al. Nod2-dependent regulation of innate and adaptive immunity in the intestinal tract. Science 307, 731–734 (2005)
Bertin, J. et al. Human CARD4 protein is a novel CED-4/Apaf-1 cell death family member that activates NF-κB. J. Biol. Chem. 274, 12955–12958 (1999)
Inohara, N. et al. Nod1, an Apaf-1-like activator of caspase-9 and nuclear factor-κB. J. Biol. Chem. 274, 14560–14567 (1999)
Faustin, B. et al. Reconstituted NALP1 inflammasome reveals two-step mechanism of caspase-1 activation. Mol. Cell 25, 713–724 (2007)
Bruey, J. M. et al. Bcl-2 and Bcl-XL regulate proinflammatory caspase-1 activation by interaction with NALP1. Cell 129, 45–56 (2007)
Conradt, B. & Horvitz, H. R. The C. elegans protein EGL-1 is required for programmed cell death and interacts with the Bcl-2-like protein CED-9. Cell 93, 519–529 (1998)
Strasser, A. The role of BH3-only proteins in the immune system. Nature Rev. Immunol. 5, 189–200 (2005)
Desagher, S. et al. Phosphorylation of bid by casein kinases I and II regulates its cleavage by caspase 8. Mol. Cell 8, 601–611 (2001)
Xavier, R. J. & Podolsky, D. K. Unravelling the pathogenesis of inflammatory bowel disease. Nature 448, 427–434 (2007)
Luo, W. et al. Bid mediates anti-apoptotic COX-2 induction through the IKKβ/NFκB pathway due to 5-MCDE exposure. Curr. Cancer Drug Targets 10, 96–106 (2010)
Wang, K., Yin, X. M., Chao, D. T., Milliman, C. L. & Korsmeyer, S. J. BID: a novel BH3 domain-only death agonist. Genes Dev. 10, 2859–2869 (1996)
Faustin, B. et al. Mechanism of Bcl-2 and Bcl-X(L) inhibition of NLRP1 inflammasome: loop domain-dependent suppression of ATP binding and oligomerization. Proc. Natl Acad. Sci. USA 106, 3935–3940 (2009)
Petros, A. M., Olejniczak, E. T. & Fesik, S. W. Structural biology of the Bcl-2 family of proteins. Biochim. Biophys. Acta 1644, 83–94 (2004)
Hasegawa, M. et al. A critical role of RICK/RIP2 polyubiquitination in Nod-induced NF-κB activation. EMBO J. 27, 373–383 (2007)
Abbott, D. W., Wilkins, A., Asara, J. M. & Cantley, L. C. The Crohn’s disease protein, NOD2, requires RIP2 in order to induce ubiquitinylation of a novel site on NEMO. Curr. Biol. 14, 2217–2227 (2004)
Perkins, N. D. Integrating cell-signalling pathways with NF-κB and IKK function. Nature Rev. Mol. Cell Biol. 8, 49–62 (2007)
Hsu, Y. M. et al. The adaptor protein CARD9 is required for innate immune responses to intracellular pathogens. Nature Immunol. 8, 198–205 (2007)
Watanabe, T. et al. Muramyl dipeptide activation of nucleotide-binding oligomerization domain 2 protects mice from experimental colitis. J. Clin. Invest. 118, 545–559 (2008)
Bertrand, M. J. et al. Cellular inhibitors of apoptosis cIAP1 and cIAP2 are required for innate immunity signaling by the pattern recognition receptors NOD1 and NOD2. Immunity 30, 789–801 (2009)
Vijay-Kumar, M. et al. Activation of toll-like receptor 3 protects against DSS-induced acute colitis. Inflamm. Bowel Dis. 13, 856–864 (2007)
Ogura, Y. et al. Nod2, a Nod1/Apaf-1 family member that is restricted to monocytes and activates NF-κB. J. Biol. Chem. 276, 4812–4818 (2001)
Boutros, M., Bras, L. P. & Huber, W. Analysis of cell-based RNAi screens. Genome Biol. 7, R66 (2006)
Zhang, J. H., Chung, T. D. & Oldenburg, K. R. A simple statistical parameter for use in evaluation and validation of high throughput screening assays. J. Biomol. Screen. 4, 67–73 (1999)
Mi, H. et al. The PANTHER database of protein families, subfamilies, functions and pathways. Nucleic Acids Res. 33, D284–D288 (2005)
Rivals, I. et al. Enrichment or depletion of a GO category within a class of genes: which test? Bioinformatics 23, 401–407 (2007)
Eisen, M. B. et al. Cluster analysis and display of genome-wide expression patterns. Proc. Natl Acad. Sci. USA 95, 14863–14868 (1998)
Saldanha, A. J. Java Treeview-extensible visualization of microarray data. Bioinformatics 20, 3246–3248 (2004)
Zhai, D. et al. Humanin binds and nullifies Bid activity by blocking its activation of Bax and Bak. J. Biol. Chem. 280, 15815–15824 (2005)
Acknowledgements
We thank A. Strasser for providing Bid−/− mice, G. Shore for BID antibodies and the McGill University high throughput/high content screening facility. We also thank D. Zhai for BID purification. This work was supported by grants from the Canadian Institutes for Health Research (CIHR-MOP 82801) and the Burroughs Wellcome Fund to M.S. M.S. is a Canadian Institutes for Health Research New Investigator. G.Y. is supported by a PDF-Fellowship from the McGill University Health Center. C.P.D. is supported by a fellowship grant from the SASS Foundation for Medical Research.
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Contributions
G.Y. and M.S. designed the research. G.Y., K.D. and M.S. performed the screen. G.Y. and M.S. analysed the data. G.Y. performed most experiments. R.G.C. performed Ni/NTA pull-down assay; P.F., R.G.C., C.P.D., D.R.G. and J.C.R. contributed new reagents/analytical tools. G.Y. and M.S. wrote the paper.
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Supplementary information
Supplementary Figures
The file contains Supplementary Figures 1-5 with legends. Supplementary Tables 1-13 were omitted when this paper first appeared online, but added on 18 May 2011. (PDF 334 kb)
Supplementary Table 1
This table shows the 225 positive regulators of NOD1 signaling identified in the NOD1 primary screen. The genes are arranged in alphabetical order. The siRNA sequences are shown in columns F and G. The Z-scores are listed in column H. (XLS 80 kb)
Supplementary Table 2
This table shows the 198 negative regulators of NOD1 signaling identified in the NOD1 primary screen. The genes are arranged in alphabetical order. The siRNA sequences are shown in columns F and G. The Z-scores are listed in column H. (XLS 71 kb)
Supplementary Table 3
This table shows the gene list comparison between “Screen 1” and “the validation screen” and identifies 200 common genes. The genes are arranged in alphabetical order. The Z-scores are listed in columns F and G. (XLS 69 kb)
Supplementary Table 4
This table shows the gene list comparison between “Screen 1” and “the TNF secondary screen” and identifies 114 common genes. The genes are arranged in alphabetical order. The Z-scores are listed in columns F and G. (XLS 51 kb)
Supplementary Table 5.
This table shows the gene list comparison between “the validation screen” and “the TNF secondary screen” identifies and 60 common genes. The genes are arranged in alphabetical order. The Z-scores are listed in columns F and G. (XLS 41 kb)
Supplementary Table 6
This table shows the classification of the NOD1 siRNA screen hits into biological processes. Genes identified from the NOD1 siRNA primary screen were classified into biological processes using the PANTHER classification system and according to GO ontology. The biological processes of positive regulators of NOD1 signaling are presented. Genes with unassigned annotations are included in the tables as ‘unclassified’. (XLS 103 kb)
Supplementary Table 7
This table shows the classification of the NOD1 siRNA screen hits into biological processes. Genes identified from the NOD1 siRNA primary screen were classified into biological processes using the PANTHER classification system and according to GO ontology. The biological processes of negative regulators of NOD1 signaling are presented. Genes with unassigned annotations are included in the tables as ‘unclassified’. (XLS 90 kb)
Supplementary Table 8
This table shows the classification of the NOD1 siRNA screen hits into molecular functions. Genes identified from the NOD1 siRNA primary screen were classified into molecular functions using the PANTHER classification system and according to GO ontology. The molecular functions of positive (regulators of NOD1 signaling are presented. Genes with unassigned annotations are included in the tables as ‘unclassified’. (XLS 71 kb)
Supplementary Table 9
This table shows the classification of the NOD1 siRNA screen hits into molecular functions. Genes identified from the NOD1 siRNA primary screen were classified into molecular functions using the PANTHER classification system and according to GO ontology. The molecular functions of negative (regulators of NOD1 signaling are presented. Genes with unassigned annotations are included in the tables as ‘unclassified’. (XLS 63 kb)
Supplementary Table 10
This table shows the validated ‘hits’ identified as enriched in immune tissues/cells. Genes that show expression enrichment in immune cells and tissues determined by Wilcoxon rank-sum test (p<0.05) are presented. (XLS 23 kb)
Supplementary Table 11
This table shows the validated ‘hits’ identified as enriched in neuronal tissues. Genes that show expression enrichment in neuronal tissues determined by Wilcoxon rank-sum test (p<0.05) are presented. (XLS 20 kb)
Supplementary Table 12
In this table the 80 tissues in the Novartis GNF1H microarray dataset are listed. Tissues clustered in duplicate are ordered from left to right on the heat map. (XLS 28 kb)
Supplementary Table 13
This table shows a list of the primers used for cloning, mutagenesis and quantitative real-time PCR experiments. (XLS 26 kb)
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Yeretssian, G., Correa, R., Doiron, K. et al. Non-apoptotic role of BID in inflammation and innate immunity. Nature 474, 96–99 (2011). https://doi.org/10.1038/nature09982
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DOI: https://doi.org/10.1038/nature09982
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