Detalle Publicación


Structure and dynamics of the second CARD of human RIG-I provide mechanistic insights into regulation of RIG-I activation

Autores: Ferrage F; Dutta K; Nistal Villan, Estanislao; Patel JR; Sánchez-Aparicio MT; De Ioannes P; Buku A; González Aseguinolaza, Gloria; García-Sastre A; Aggarwal AK
Título de la revista: STRUCTURE
ISSN: 0969-2126
Volumen: 20
Número: 12
Páginas: 2048-2061
Fecha de publicación: 2012
RIG-I is a cytosolic sensor of viral RNA, comprised of two N-terminal CARDs followed by helicase and C-terminal regulatory domains (helicase-CTD). Viral RNA binds to the helicase-CTD and "exposes" the CARDs for downstream signaling. The role of the second CARD (CARD2) is essential as RIG-I activation requires dephosphorylation of Thr170 followed by ubiquitination at Lys172. Here, we present the solution structure and dynamics of human RIG-I CARD2. Surprisingly, we find that Thr170 is mostly buried. Parallel studies on the phosphomimetic T170E mutant suggest that the loss of function upon Thr170 phosphorylation is likely associated with changes in the CARD1-CARD2 interface that may prevent Lys172 ubiquitination and/or binding to free K63-linked polyubiquitin. We also demonstrate a strong interaction between CARD2 and the helicase-CTD, and show that mutations at the interface result in constitutive activation of RIG-I. Collectively, our data suggests a close interplay between phosphorylation, ubiquitination, and activation of human RIG-I, all mediated by CARD2.