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|Title:||Structural insights into RISC assembly facilitated by dsRNA-binding domains of human RNA helicase A (DHX9)||Authors:||Fu, Q.
|Issue Date:||Mar-2013||Citation:||Fu, Q., Yuan, Y.A. (2013-03). Structural insights into RISC assembly facilitated by dsRNA-binding domains of human RNA helicase A (DHX9). Nucleic Acids Research 41 (5) : 3457-3470. ScholarBank@NUS Repository. https://doi.org/10.1093/nar/gkt042||Abstract:||ABSTRACT Intensive research interest has focused on small RNA-processing machinery and the RNA-induced silencing complex (RISC), key cellular machines in RNAi pathways. However, the structural mechanism regarding RISC assembly, the primary step linking small RNA processing and RNA-mediated gene silencing, is largely unknown. Human RNA helicase A (DHX9) was reported to function as an RISCloading factor, and such function is mediated mainly by its dsRNA-binding domains (dsRBDs). Here, we report the crystal structures of human RNA helicase A (RHA) dsRBD1 and dsRBD2 domains in complex with dsRNAs, respectively. Structural analysis not only reveals higher siRNA duplex-binding affinity displayed by dsRBD1, but also identifies a crystallographic dsRBD1 pair of physiological significance in cooperatively recognizing dsRNAs. Structural observations are further validated by isothermal titration calorimetric (ITC) assay. Moreover, co-immunoprecipitation (co-IP) assay coupled with mutagenesis demonstrated that both dsRBDs are required for RISC association, and such association is mediated by dsRNA. Hence, our structural and functional efforts have revealed a potential working model for siRNA recognition by RHA tandem dsRBDs, and together they provide direct structural insights into RISC assembly facilitated by RHA. © The Author(s) 2013.||Source Title:||Nucleic Acids Research||URI:||http://scholarbank.nus.edu.sg/handle/10635/101762||ISSN:||03051048||DOI:||10.1093/nar/gkt042|
|Appears in Collections:||Staff Publications|
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