The Protein ATG16L1 Suppresses Inflammatory Cytokines Induced by the Intracellular Sensors Nod1 and Nod2 in an Autophagy-Independent Manner

The peptidoglycan sensor Nod2 and the autophagy protein ATG16L1 have been linked to Crohn’s disease (CD). Although Nod2 and the related sensor, Nod1, direct ATG16L1 to initiate anti-bacterial autophagy, whether ATG16L1 affects Nod-driven inflammation has not been examined. Here, we uncover an unanticipated autophagy-independent role for ATG16L1 in negatively regulating Nod-driven inflammatory responses. Knockdown of ATG16L1 expression, but not that of ATG5 or ATG9a, specifically enhanced Nod-driven cytokine production. In addition, autophagy-incompetent truncated forms of ATG16L1 regulated Nod-driven cytokine responses. Mechanistically, we demonstrated that ATG16L1 interfered with poly-ubiquitination of the Rip2 adaptor and recruitment of Rip2 into large signaling complexes. The CD-associated allele of ATG16L1 was impaired in its ability to regulate Nod-driven inflammatory responses. Overall, these results suggest that ATG16L1 is critical for Nod-dependent regulation of cytokine responses and that disruption of this Nod1- or Nod2-ATG16L1 signaling axis could contribute to the chronic inflammation associated with CD. •ATG16L1 suppresses Nod1- and Nod2-driven cytokine responses•ATG16L1’s regulatory function is independent of its role in autophagosome formation•ATG16L1 negatively regulates Nod1 and Nod2 signaling via Rip2 activation•Crohn’s-disease-associated ATG16L1 allele is defective in Nod1 and Nod2 regulation