ciap12 are direct e3 ligases conjugating diverse types of ubiquitin chains to receptor interacting proteins kinases 1 to 4 (rip1–4)ciap12直接e3连接接合不同类型的泛素链受体相互作用蛋白激酶1到4(rip1-4).pdfVIP

cIAP1/2 Are Direct E3 Ligases Conjugating Diverse Types of Ubiquitin Chains to Receptor Interacting Proteins Kinases 1 to 4 (RIP1–4) Mathieu J. M. Bertrand1,2*, Saskia Lippens1,2, An Staes3,4, Barbara Gilbert1,2, Ria Roelandt1,2, Jelle De Medts1,2, Kris Gevaert3,4, Wim Declercq1,2., Peter Vandenabeele1,2. 1 Department for Molecular Biomedical Research, VIB, Ghent, Belgium, 2 Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium, 3 Department of Medical Protein Research, VIB, Ghent, Belgium, 4 Department of Biochemistry, Ghent University, Ghent, Belgium Abstract The RIP kinases have emerged as essential mediators of cellular stress that integrate both extracellular stimuli emanat- ing from various cell-surface receptors and signals coming from intracellular pattern recognition receptors. The molecular mechanisms regulating the ability of the RIP proteins to transduce the stress signals remain poorly understood, but seem to rely only partially on their kinase activities. Recent studies on RIP1 and RIP2 have highlighted the importance of ubiquitination as a key process regulating their capacity to activate downstream signaling pathways. In this study, we found that XIAP, cIAP1 and cIAP2 not only directly bind to RIP1 and RIP2 but also to RIP3 and RIP4. We show that cIAP1 and cIAP2 are direct E3 ubiquitin ligases for all four RIP proteins and that cIAP1 is capable of conjugating the RIPs with diverse types of ubiquitin chains, including linear chains. Consistently, we show that repressing cIAP1/2 levels affects the activation of NF-kB that is dependent on RIP1, -2, -3 and -4. Finally, we identified Lys51 and Lys145 of RIP4 as two critical residues for cIAP1- mediated ubiquitination and NF-kB activation. Citation: Bertrand MJM, Lippens S, Staes A, Gilbert B, Roelandt R, et al. (2011) cIAP1/2 Are Direct E3 Ligases Conjugating Diverse Types