Ubiquitination of ATF6 by disease-associated RNF186 promotes the innate receptor-induced unfolded protein response.

Properly balancing microbial responses by the innate immune system through pattern recognition receptors (PRRs) is critical for intestinal immune homeostasis. Ring finger protein 186 (RNF186) genetic variants are associated with inflammatory bowel disease (IBD). However, functions for the E3 ubiquitin ligase RNF186 are incompletely defined. We found that upon stimulation of the PRR nucleotide-binding oligomerization domain containing 2 (NOD2) in human macrophages, RNF186 localized to the ER, formed a complex with ER stress sensors, ubiquitinated the ER stress sensor activating transcription factor 6 (ATF6), and promoted the unfolded protein response (UPR). These events, in turn, led to downstream signaling, cytokine secretion, and antimicrobial pathway induction. Importantly, RNF186-mediated ubiquitination of K152 on ATF6 was required for these outcomes, highlighting a key role for ATF6 ubiquitination in PRR-initiated functions. Human macrophages transfected with the rare RNF186-A64T IBD risk variant and macrophages from common rs6426833 RNF186 IBD risk carriers demonstrated reduced NOD2-induced outcomes, which were restored by rescuing UPR signaling. Mice deficient in RNF186 or ATF6 demonstrated a reduced UPR in colonic tissues, increased weight loss, and less effective clearance of bacteria with dextran sodium sulfate-induced injury and upon oral challenge with Salmonella Typhimurium. Therefore, we identified that RNF186 was required for PRR-induced, UPR-associated signaling leading to key macrophage functions; defined that RNF186-mediated ubiquitination of ATF6 was essential for these functions; and elucidated how RNF186 IBD risk variants modulated these outcomes.

[1]  Chenhui Wang,et al.  RNF186 regulates EFNB1 (ephrin B1)-EPHB2-induced autophagy in the colonic epithelial cells for the maintenance of intestinal homeostasis , 2020, Autophagy.

[2]  Huabing Zhang,et al.  The role and mechanism of action of RNF186 in colorectal cancer through negative regulation of NF-κB. , 2020, Cellular signalling.

[3]  C. Abraham,et al.  TNFSF15 Promotes Antimicrobial Pathways in Human Macrophages and These Are Modulated by TNFSF15 Disease-Risk Variants , 2020, Cellular and molecular gastroenterology and hepatology.

[4]  C. Abraham,et al.  Myeloid Cell Expression of LACC1 is Required for Bacterial Clearance and Control of Intestinal Inflammation. , 2020, Gastroenterology.

[5]  A. Kaser,et al.  Activating Transcription Factor 6 Mediates Inflammatory Signals in Intestinal Epithelial Cells Upon Endoplasmic Reticulum Stress. , 2020, Gastroenterology.

[6]  C. Abraham,et al.  LACC1 Required for NOD2-Induced, ER Stress-Mediated Innate Immune Outcomes in Human Macrophages and LACC1 Risk Variants Modulate These Outcomes. , 2019, Cell reports.

[7]  Bill B. Chen,et al.  The RING-type E3 ligase RNF186 ubiquitinates Sestrin-2 and thereby controls nutrient sensing , 2019, The Journal of Biological Chemistry.

[8]  Huabing Zhang,et al.  RNF186 impairs insulin sensitivity by inducing ER stress in mouse primary hepatocytes. , 2018, Cellular signalling.

[9]  C. Hetz,et al.  The Unfolded Protein Response and Cell Fate Control. , 2017, Molecular cell.

[10]  R. Xavier,et al.  Erratum: E3 Ubiquitin ligase ZNRF4 negatively regulates NOD2 signalling and induces tolerance to MDP , 2017, Nature Communications.

[11]  R. Xavier,et al.  E3 Ubiquitin ligase ZNRF4 negatively regulates NOD2 signalling and induces tolerance to MDP , 2017, Nature Communications.

[12]  C. Abraham,et al.  Human LACC1 increases innate receptor-induced responses and a LACC1 disease-risk variant modulates these outcomes , 2017, Nature Communications.

[13]  C. Abraham,et al.  An inflammatory bowel disease–risk variant in INAVA decreases pattern recognition receptor–induced outcomes , 2017, The Journal of clinical investigation.

[14]  M. Peter,et al.  Translocon component Sec62 acts in endoplasmic reticulum turnover during stress recovery , 2016, Nature Cell Biology.

[15]  K. Takeda,et al.  Regulation of intestinal homeostasis by the ulcerative colitis-associated gene RNF186 , 2016, Mucosal Immunology.

[16]  Randal J. Kaufman,et al.  The unfolded protein response in immunity and inflammation , 2016, Nature Reviews Immunology.

[17]  Denise N. Bronner,et al.  Endoplasmic Reticulum Stress Activates the Inflammasome via NLRP3- and Caspase-2-Driven Mitochondrial Damage. , 2015, Immunity.

[18]  Chad D Williamson,et al.  Isolation of Endoplasmic Reticulum, Mitochondria, and Mitochondria‐Associated Membrane and Detergent Resistant Membrane Fractions from Transfected Cells and from Human Cytomegalovirus‐Infected Primary Fibroblasts , 2015, Current protocols in cell biology.

[19]  Z. Ling,et al.  The Endoplasmic Reticulum Stress Sensor IRE1α in Intestinal Epithelial Cells Is Essential for Protecting against Colitis* , 2015, The Journal of Biological Chemistry.

[20]  L. Glimcher,et al.  Endoplasmic reticulum stress in immunity. , 2015, Annual review of immunology.

[21]  C. Abraham,et al.  Activation of pattern recognition receptors up-regulates metallothioneins, thereby increasing intracellular accumulation of zinc, autophagy, and bacterial clearance by macrophages. , 2014, Gastroenterology.

[22]  Siqi Liu,et al.  Ubiquitination of Inositol-requiring Enzyme 1 (IRE1) by the E3 Ligase CHIP Mediates the IRE1/TRAF2/JNK Pathway* , 2014, The Journal of Biological Chemistry.

[23]  C. Abraham,et al.  A TNFSF15 disease-risk polymorphism increases pattern-recognition receptor-induced signaling through caspase-8–induced IL-1 , 2014, Proceedings of the National Academy of Sciences.

[24]  M. Silverberg,et al.  Variants in nicotinamide adenine dinucleotide phosphate oxidase complex components determine susceptibility to very early onset inflammatory bowel disease. , 2014, Gastroenterology.

[25]  J. Rao,et al.  ATF6 Mediates a Pro‐Inflammatory Synergy Between ER Stress and TLR Activation in the Pathogenesis of Liver Ischemia‐Reperfusion Injury , 2014, American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons.

[26]  Judy H. Cho,et al.  Pattern recognition receptor signaling in human dendritic cells is enhanced by ICOS ligand and modulated by the Crohn's disease ICOSLG risk allele. , 2014, Immunity.

[27]  C. Abraham,et al.  NOD2 Regulates CXCR3-Dependent CD8+ T Cell Accumulation in Intestinal Tissues with Acute Injury , 2014, The Journal of Immunology.

[28]  Yan Li,et al.  A novel RING finger E3 ligase RNF186 regulate ER stress-mediated apoptosis through interaction with BNip1. , 2013, Cellular signalling.

[29]  C. Abraham,et al.  NF-κB1 Inhibits NOD2-Induced Cytokine Secretion through ATF3-Dependent Mechanisms , 2013, Molecular and Cellular Biology.

[30]  H. Perlman,et al.  Toll‐like receptor‐mediated IRE1α activation as a therapeutic target for inflammatory arthritis , 2013, The EMBO journal.

[31]  Judy H. Cho,et al.  Deep Resequencing of GWAS Loci Identifies Rare Variants in CARD9, IL23R and RNF186 That Are Associated with Ulcerative Colitis , 2013, PLoS genetics.

[32]  Daniel R. Richards,et al.  Genomic responses in mouse models poorly mimic human inflammatory diseases , 2013, Proceedings of the National Academy of Sciences.

[33]  C. Abraham,et al.  Nod2-induced autocrine interleukin-1 alters signaling by ERK and p38 to differentially regulate secretion of inflammatory cytokines. , 2012, Gastroenterology.

[34]  G. Barton,et al.  Faculty Opinions recommendation of NLRC4-driven production of IL-1β discriminates between pathogenic and commensal bacteria and promotes host intestinal defense. , 2012 .

[35]  C. Hetz.  The unfolded protein response: controlling cell fate decisions under ER stress and beyond , 2012, Nature Reviews Molecular Cell Biology.

[36]  P. Walter,et al.  The Unfolded Protein Response: From Stress Pathway to Homeostatic Regulation , 2011, Science.

[37]  Clara Abraham,et al.  Interactions between the host innate immune system and microbes in inflammatory bowel disease. , 2011, Gastroenterology.

[38]  J. Achkar,et al.  ATG16L1 and NOD2 interact in an autophagy-dependent antibacterial pathway implicated in Crohn's disease pathogenesis. , 2010, Gastroenterology.

[39]  A. Kaser,et al.  Endoplasmic reticulum stress: implications for inflammatory bowel disease pathogenesis , 2010, Current opinion in gastroenterology.

[40]  Xi Chen,et al.  TLR activation of the transcription factor XBP1 regulates innate immune responses in macrophages , 2010, Nature Immunology.

[41]  M. Permutt,et al.  Wolfram syndrome 1 gene negatively regulates ER stress signaling in rodent and human cells. , 2010, The Journal of clinical investigation.

[42]  A. Towbin,et al.  Chronic granulomatous disease , 2010, Pediatric Radiology.

[43]  H. Tilg,et al.  XBP1 Links ER Stress to Intestinal Inflammation and Confers Genetic Risk for Human Inflammatory Bowel Disease , 2008, Cell.

[44]  Donna D. Zhang,et al.  Synoviolin promotes IRE1 ubiquitination and degradation in synovial fibroblasts from mice with collagen‐induced arthritis , 2008, EMBO reports.

[45]  Ping-Chang Yang,et al.  Mast cells play a crucial role in Staphylococcus aureus peptidoglycan-induced diarrhea. , 2007, The American journal of pathology.

[46]  P. Walter,et al.  Signal integration in the endoplasmic reticulum unfolded protein response , 2007, Nature Reviews Molecular Cell Biology.

[47]  J. Orenstein,et al.  Human intestinal macrophages display profound inflammatory anergy despite avid phagocytic and bacteriocidal activity. , 2005, The Journal of clinical investigation.

[48]  Laurent Beaugerie,et al.  High prevalence of adherent-invasive Escherichia coli associated with ileal mucosa in Crohn's disease. , 2004, Gastroenterology.

[49]  Amy S. Lee,et al.  Endoplasmic reticulum stress triggers an acute proteasome‐dependent degradation of ATF6 , 2004, Journal of cellular biochemistry.

[50]  Ron Prywes,et al.  The Luminal Domain of ATF6 Senses Endoplasmic Reticulum (ER) Stress and Causes Translocation of ATF6 from the ER to the Golgi* , 2002, The Journal of Biological Chemistry.

[51]  D. Nicolae,et al.  A frameshift mutation in NOD2 associated with susceptibility to Crohn's disease , 2001, Nature.

[52]  Judy H. Cho,et al.  Increased sensitivity to dextran sodium sulfate colitis in IRE1beta-deficient mice. , 2001, The Journal of clinical investigation.

[53]  Judy H. Cho,et al.  Materials for : Host-microbe interactions shape genetic risk for inflammatory bowel disease , 2012 .

[54]  E. Szigethy,et al.  Inflammatory bowel disease. , 2011, Pediatric clinics of North America.

[55]  D. Philpott,et al.  Nod1 and Nod2 direct autophagy by recruiting ATG16L1 to the plasma membrane at the site of bacterial entry , 2010, Nature Immunology.

[56]  Simmie L. Foster,et al.  Gene-specific control of inflammation by TLR-induced chromatin modifications , 2008, Nature.

[57]  M. Dinauer,et al.  Phenotype of mice and macrophages deficient in both phagocyte oxidase and inducible nitric oxide synthase. , 1999, Immunity.