Journal article
Characterization of the TRBP domain required for Dicer interaction and function in RNA interference
SM Daniels, CE Melendez-Peña, RJ Scarborough, A Daher, HS Christensen, M El Far, DFJ Purcell, S Lainé, A Gatignol
BMC Molecular Biology | Published : 2009
Open access
Abstract
Background: Dicer, Ago2 and TRBP are the minimum components of the human RNA-induced silencing complex (RISC). While Dicer and Ago2 are RNases, TRBP is the double-stranded RNA binding protein (dsRBP) that loads small interfering RNA into the RISC. TRBP binds directly to Dicer through its C-terminal domain. Results: We show that the TRBP binding site in Diceris a 165 amino acid (aa) region located between the ATPase and the helicase domains. The binding site in TRBP is a 69 aa domain, called C4, located at the C-terminal end of TRBP. The TRBP1 and TRBP2 isoforms, but not TRBPs lacking the C4 site (TRBPsΔC4), co-immunoprecipitated with Dicer. The C4 domain is therefore necessary to bind Dicer,..
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Awarded by Canadian Institutes of Health Research (CIHR)
Funding Acknowledgements
We are very grateful to Dr. W. Filipowicz for the generous gift of the Dicer antibodies. We also thank Dr. Laurence Lejeune for help in FACS analysis, Dr. Judith Lacoste for help in confocal microscopy and Guerline Clerzius and Jean-Francois Gelinas for comments on the manuscript. This work was supported by grant HOP38112 and HOP93434 from the Canadian Institutes of Health Research (CIHR) (to AG). RS is supported by a Frederick Banting and Charles Best Canada Graduate Scholarships, Master's award from CIHR. SL was supported by a post-doctoral fellowship from CIHR. AG was a recipient of a Hugh and Helen McPherson Memorial Salary Award.