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Expanded RNA-binding activities of mammalian Argonaute 2.

Research paper by Grace S GS Tan, Barry G BG Garchow, Xuhang X Liu, Jennifer J Yeung, John P JP Morris, Trinna L TL Cuellar, Michael T MT McManus, Marianthi M Kiriakidou

Indexed on: 08 Oct '09Published on: 08 Oct '09Published in: Nucleic acids research



Abstract

Mammalian Argonaute 2 (Ago2) protein associates with microRNAs (miRNAs) or small interfering RNAs (siRNAs) forming RNA-induced silencing complexes (RISCs/miRNPs). In the present work, we characterize the RNA-binding and nucleolytic activity of recombinant mouse Ago2. Our studies show that recombinant mouse Ago2 binds efficiently to miRNAs forming active RISC. Surprisingly, we find that recombinant mouse Ago2 forms active RISC using pre-miRNAs or long unstructured single stranded RNAs as guides. Furthermore, we demonstrate that, in vivo, endogenous human Ago2 binds directly to pre-miRNAs independently of Dicer, and that Ago2:pre-miRNA complexes are found both in the cytoplasm and in the nucleus of human cells.