J Biol Chem. 2011 Aug 12;286(32):28574-83. Epub 2011 Jun 20.

Retinoic acid-induced gene-I (RIG-I) associates with nucleotide-binding oligomerization domain-2 (NOD2) to negatively regulate inflammatory signaling.External

Morosky, S. A., Zhu, J., Mukherjee, A., Sarkar, S. N., Coyne, C. B.,
--- - Department of Microbiology and Molecular Genetics, University of Pittsburgh Cancer Institute, University of Pittsburgh, Pittsburgh, Pennsylvania 15219, USA.
Cytoplasmic caspase recruiting domain (CARD)-containing molecules often function in the induction of potent antimicrobial responses in order to protect mammalian cells from invading pathogens. Retinoic acid-induced gene-I (RIG-I) and nucleotide binding oligomerization domain 2 (NOD2) serve as key factors in the detection of viral and bacterial pathogens, and in the subsequent initiation of innate immune signals to combat infection. RIG-I and NOD2 share striking similarities in their cellular localization, both localize to membrane ruffles in non-polarized epithelial cells and both exhibit a close association with the junctional complex of polarized epithelia. Here we show that RIG-I and NOD2 not only colocalize to cellular ruffles and cell-cell junctions, but that they also form a direct interaction that is mediated by the CARDs of RIG-I and multiple regions of NOD2. Moreover, we show that RIG-I negatively regulates ligand-induced nuclear factor-kappaB (NF-kappaB) signaling mediated by NOD2, and that NOD2 negatively regulates type I interferon induction by RIG-I. We also show that the three main Crohn disease-associated mutants of NOD2 (1007fs, R702W, G908R) form an interaction with RIG-I and negatively regulate its signaling to a greater extent than wild-type NOD2. Our results show that in addition to their role in innate immune recognition, RIG-I and NOD2 form a direct interaction at actin-enriched sites within cells and suggest that this interaction may impact RIG-I- and NOD2-dependent innate immune signaling.
PMID: 21690088External
Arrow2 In vitro interaction Arrow2 In vivo interaction Arrow2 Characterization Arrow2 Functional role Arrow2 top
In vitro interaction
  DD1 DD2 Reference
Family DD1 DD2 Method Species Region Expression Species Region Expression
CARD NOD2 Link RIG-1 His Fusion protein pull-down Not specified 1-301 E.coli Not specified 1-205 E.coli 21690088
(Link: click this icon to show interactions only between the two corresponding DDs)
Arrow2 In vitro interaction Arrow2 In vivo interaction Arrow2 Characterization Arrow2 Functional role Arrow2 top
In vivo interaction
  Endogenous
expression
Overexpression DD1 DD2 Reference
Family DD1 DD2 Method Species Region Species Region
CARD NOD2 Link RIG-1 Co-Immunofluorescence staining HT29 21690088
CARD NOD2 Link RIG-1 Co-Immunofluorescence staining HEK293 Not specified Not specified Not specified Not specified 21690088
CARD NOD2 Link RIG-1 Co-immunoprecipitation HEK293 Not specified Full length Not specified Full length 21690088
CARD NOD2 Link RIG-1 Co-immunoprecipitation HEK293 Not specified 1-301 Not specified Full length 21690088
CARD NOD2 Link RIG-1 Co-immunoprecipitation HEK293 Not specified 1-301 Not specified 1-225 21690088
CARD NOD2 Link RIG-1 Co-immunoprecipitation HEK293 Not specified Full length Not specified 1-225 21690088
(Link: click this icon to show interactions only between the two corresponding DDs)
Arrow2 In vitro interaction Arrow2 In vivo interaction Arrow2 Characterization Arrow2 Functional role Arrow2 top
Functional Role
Family DD1 DD2 Method Death-related Death-unrelated Reference
CARD NOD2 Link RIG-1 Reporter gene assay NF-kB activation block, IFN-b activation block 21690088
(Link: click this icon to show interactions only between the two corresponding DDs)