Yang, YibinYin, Catherine C.Pandey, Amit K.Abbott, DerekSassetti, Christopher MKelliher, Michelle A.2022-08-232022-08-232007-12-142009-04-07J Biol Chem. 2007 Dec 14;282(50):36223-9. Epub 2007 Oct 18. <a href="http://dx.doi.org/10.1074/jbc.M703079200">Link to article on publisher's website</a>0021-925810.1074/jbc.M7030792001794723617947236https://hdl.handle.net/20.500.14038/33041The Rip2 kinase contains a caspase recruitment domain and has been implicated in the activation of the transcriptional factor NF-kappaB downstream of Toll-like receptors, Nod-like receptors, and the T cell receptor. Although Rip2 has been linked to Nod signaling, how Nod-Rip2 proteins mediate NF-kappaB activation has remained unclear. We find Rip2 required for Nod2-mediated NF-kappaB activation and to a lesser extent mitogen-activated protein kinase activation. We demonstrate that Rip2 and IkappaB kinase-gamma become stably polyubiquitinated upon treatment of cells with the NOD2 ligand, muramyl dipeptide. We also demonstrate a requirement for the E2-conjugating enzyme Ubc13, the E3 ubiquitin ligase Traf6, and the ubiquitin-activated kinase Tak1 in Nod2-mediated NF-kappaB activation. Rip2 polyubiquitination is also stimulated when macrophages are infected with live Mycobacterium tuberculosis but not when infected with heat-killed bacteria. Consistent with our data linking Rip2 to NOD and not Toll-like receptor signaling, M. tuberculosis-induced Rip2 polyubiquitination appears MyD88-independent. Collectively, these data reveal that the NOD2 pathway is ubiquitin-regulated and that Rip2 employs a ubiquitin-dependent mechanism to achieve NF-kappaB activation.en-USJournal Articlehttps://escholarship.umassmed.edu/gsbs_sp/1590814159gsbs_sp/1590