TY - JOUR
T1 - The kinase-activity of Rip2 determines its stability and consequently Nod1 - and Nod2-mediated immune responses
AU - Nembrini, Chiara
AU - Kisielow, Jan
AU - Shamshiev, Abdijapar T.
AU - Tortola, Luigi
AU - Coyle, Anthony J.
AU - Kopf, Manfred
AU - Marsland, Benjamin J.
PY - 2009/7/17
Y1 - 2009/7/17
N2 - Rip2 (RICK, CARD3) has been identified as a key effector molecule downstream of the pattern recognition receptors, Nod1 and Nod2; however, its mechanism of action remains to be elucidated. In particular, it is unclear whether its kinase activity is required for signaling or for maintaining protein stability. We have investigated the expression level of different retrovirally expressed kinase-dead Rip2 mutants and the role of Rip2 kinase activity in the signaling events that follow Nod1 and Nod2 stimulation. We show that in primary cells expressing kinase-inactive Rip2, protein levels were severely compromised, and stability could not be reconstituted by the addition of a phospho-mimetic mutation in its autophosphorylation site. Consequently, inflammatory cytokine production in response to Nod1 and Nod2 ligands was abrogated both in vitro and in vivo in the absence of Rip2 kinase activity. Our results highlight the central role that Rip2 kinase activity plays in conferring stability to the protein and thus in the preservation of Nod1- and Nod2-mediated innate immune responses.
AB - Rip2 (RICK, CARD3) has been identified as a key effector molecule downstream of the pattern recognition receptors, Nod1 and Nod2; however, its mechanism of action remains to be elucidated. In particular, it is unclear whether its kinase activity is required for signaling or for maintaining protein stability. We have investigated the expression level of different retrovirally expressed kinase-dead Rip2 mutants and the role of Rip2 kinase activity in the signaling events that follow Nod1 and Nod2 stimulation. We show that in primary cells expressing kinase-inactive Rip2, protein levels were severely compromised, and stability could not be reconstituted by the addition of a phospho-mimetic mutation in its autophosphorylation site. Consequently, inflammatory cytokine production in response to Nod1 and Nod2 ligands was abrogated both in vitro and in vivo in the absence of Rip2 kinase activity. Our results highlight the central role that Rip2 kinase activity plays in conferring stability to the protein and thus in the preservation of Nod1- and Nod2-mediated innate immune responses.
UR - http://www.scopus.com/inward/record.url?scp=67749095245&partnerID=8YFLogxK
U2 - 10.1074/jbc.M109.006353
DO - 10.1074/jbc.M109.006353
M3 - Article
C2 - 19473975
AN - SCOPUS:67749095245
VL - 284
SP - 19183
EP - 19188
JO - The Journal of Biological Chemistry
JF - The Journal of Biological Chemistry
SN - 1083-351X
IS - 29
ER -