期刊
NUCLEIC ACIDS RESEARCH
卷 37, 期 22, 页码 7533-7545出版社
OXFORD UNIV PRESS
DOI: 10.1093/nar/gkp812
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资金
- National Institutes of Health/National Institute of Allergy and Infectious Diseases [K08AI063030]
- University of Pennsylvania
- National Institutes of Health/National Institute of Arthritis and Musculoskeletal and Skin Diseases [T32-AR007442-22]
- National Institutes of Health/National Institute of General Medical Sciences [R01GM080783]
- National Institutes of Health and University of Pennsylvania
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.
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