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Mol Cell. 2015 Sep 3;59(5):807-18. doi: 10.1016/j.molcel.2015.07.006. Epub 2015 Aug 6.

TAF11 Assembles the RISC Loading Complex to Enhance RNAi Efficiency.

Author information

1
Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA.
2
Department of Biological Sciences, University of Tokyo, Tokyo 113-0032, Japan.
3
Departments of Pharmacology and Neuroscience, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA.
4
Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA; International Institute of Integrated Sleep Medicine, University of Tsukuba, Tsukuba 305-8575, Japan. Electronic address: qinghua.liu@utsouthwestern.edu.

Abstract

Assembly of the RNA-induced silencing complex (RISC) requires formation of the RISC loading complex (RLC), which contains the Dicer-2 (Dcr-2)-R2D2 complex and recruits duplex siRNA to Ago2 in Drosophila melanogaster. However, the precise composition and action mechanism of Drosophila RLC remain unclear. Here we identified the missing factor of RLC as TATA-binding protein-associated factor 11 (TAF11) by genetic screen. Although it is an annotated nuclear transcription factor, we found that TAF11 also associated with Dcr-2/R2D2 and localized to cytoplasmic D2 bodies. Consistent with defective RLC assembly in taf11(-/-) ovary extract, we reconstituted the RLC in vitro using the recombinant Dcr-2-R2D2 complex, TAF11, and duplex siRNA. Furthermore, we showed that TAF11 tetramer facilitates Dcr-2-R2D2 tetramerization to enhance siRNA binding and RISC loading activities. Together, our genetic and biochemical studies define the molecular nature of the Drosophila RLC and elucidate a cytoplasmic function of TAF11 in organizing RLC assembly to enhance RNAi efficiency.

PMID:
26257286
PMCID:
PMC4560963
DOI:
10.1016/j.molcel.2015.07.006
[Indexed for MEDLINE]
Free PMC Article

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