RNA silencing is a potent antiviral mechanism in plants and animals.As a counter-defense,many viruses studied to date encode one or more viral suppressors of RNA silencing (VSR).In the latter case,how different VSRs encoded by a virus function in silencing remains to be fully understood.We previously showed that the nonstructural protein Pns10 of a Phytoreovirus,Rice dwarf virus (RDV),functions as a VSR.Here we present evidence that another nonstructural protein,Pns 11,also functions as a VSR.While Pns10 was localized in the cytoplasm,Pns11 was localized both in the nucleus and chloroplasts.Pns11 has two bipartite nuclear localization signals (NLSs),which were required for nuclear as well as chloroplastic localization.The NLSs were also required for the silencing activities of Pns11.This is the first report that multiple VSRs encoded by a virus are localized in different subcellular compartments,and that a viral protein can be targeted to both the nucleus and chloroplast.These findings may have broad significance in studying the subcellular targeting of VSRs and other viral proteins in viral-host interactions.