5N8M

Structure of TRBP dsRBD 1 and 2 in complex with a 19 bp siRNA (Complex A)

Summary for 5N8M

• Entry DOI: 10.2210/pdb5n8m/pdb
• NMR Information: BMRB: 34104
• Descriptor: RISC-loading complex subunit TARBP2, RNA (5'-R(*UP*UP*AP*AP*UP*UP*AP*UP*CP*UP*AP*UP*UP*CP*CP*GP*UP*AP*CP*UP*U)-3'), RNA (5'-R(*GP*UP*AP*CP*GP*GP*AP*AP*UP*AP*GP*AP*UP*AP*AP*UP*UP*AP*AP*UP*U)-3') (3 entities in total)
• Functional Keywords: trbp, sirna, protein-rna complex, rnai, rna binding protein
• Biological source: Homo sapiens (Human); More
• Total number of polymer chains: 3
• Total formula weight: 36294.22

Authors

Masliah, G.,Maris, C.,Allain, H.-T.F. (deposition date: 2017-02-23, release date: 2018-02-21, Last modification date: 2024-05-15)

Primary citation

Masliah, G.,Maris, C.,Konig, S.L.,Yulikov, M.,Aeschimann, F.,Malinowska, A.L.,Mabille, J.,Weiler, J.,Holla, A.,Hunziker, J.,Meisner-Kober, N.,Schuler, B.,Jeschke, G.,Allain, F.H.
Structural basis of siRNA recognition by TRBP double-stranded RNA binding domains.
EMBO J., 37:-, 2018

PubMed Abstract: The accurate cleavage of pre-micro(mi)RNAs by Dicer and mi/siRNA guide strand selection are important steps in forming the RNA-induced silencing complex (RISC). The role of Dicer binding partner TRBP in these processes remains poorly understood. Here, we solved the solution structure of the two N-terminal dsRNA binding domains (dsRBDs) of TRBP in complex with a functionally asymmetric siRNA using NMR, EPR, and single-molecule spectroscopy. We find that siRNA recognition by the dsRBDs is not sequence-specific but rather depends on the RNA shape. The two dsRBDs can swap their binding sites, giving rise to two equally populated, pseudo-symmetrical complexes, showing that TRBP is not a primary sensor of siRNA asymmetry. Using our structure to model a Dicer-TRBP-siRNA ternary complex, we show that TRBP's dsRBDs and Dicer's RNase III domains bind a canonical 19 base pair siRNA on opposite sides, supporting a mechanism whereby TRBP influences Dicer-mediated cleavage accuracy by binding the dsRNA region of the pre-miRNA during Dicer cleavage.

PubMed: 29449323
DOI: 10.15252/embj.201797089

Experimental method

SOLUTION NMR