viral protein inhibits risc activity by argonaute binding through conserved wggw motifs病毒蛋白抑制risc活动通过守恒wggw图案argonaute绑定.pdfVIP

Viral Protein Inhibits RISC Activity by Argonaute Binding through Conserved WG/GW Motifs 1. ´ ´ 2 . ´ 1 ´ ´ 1 ´ ´ 2,3 Ana Giner , Lorant Lakatos * , Meritxell Garcıa-Chapa , Juan Jose Lopez-Moya *, Jozsef Burgyan * ¨ ¨ 1 Centre for Research in Agricultural Genomics, CRAG, CSIC-IRTA-UAB, Barcelona, Spain, 2 Agricultural Biotechnology Centre, Godollo˝ , Hungary, 3 Instituto di Virologia Vegetale, Torino, Italy Abstract RNA silencing is an evolutionarily conserved sequence-specific gene-inactivation system that also functions as an antiviral mechanism in higher plants and insects. To overcome antiviral RNA silencing, viruses express silencing-suppressor proteins. These viral proteins can target one or more key points in the silencing machinery. Here we show that in Sweet potato mild mottle virus (SPMMV, type member of the Ipomovirus genus, family Potyviridae), the role of silencing suppressor is played by the P1 protein (the largest serine protease among all known potyvirids) despite the presence in its genome of an HC-Pro protein, which, in potyviruses, acts as the suppressor. Using in vivo studies we have demonstrated that SPMMV P1 inhibits si/miRNA-programmed RISC activity. Inhibition of RISC activity occurs by binding P1 to mature high molecular weight RISC, as we have shown by immunoprecipitation. Our results revealed that P1 targets Argonaute1 (AGO1), the catalytic unit of RISC, and that suppressor/binding activities are localized at the N-terminal half of P1. I