TRBP ensures efficient Dicer processing of precursor microRNA in RNA-crowded environments

Mohamed Fareh, Kyu-Hyeon Yeom, Anna C. Haagsma, Sweeny Chauhan, Inha Heo and Chirlmin Joo ()
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Mohamed Fareh: Kavli Institute of NanoScience, Delft University of Technology
Kyu-Hyeon Yeom: Kavli Institute of NanoScience, Delft University of Technology
Anna C. Haagsma: Kavli Institute of NanoScience, Delft University of Technology
Sweeny Chauhan: Kavli Institute of NanoScience, Delft University of Technology
Inha Heo: Kavli Institute of NanoScience, Delft University of Technology
Chirlmin Joo: Kavli Institute of NanoScience, Delft University of Technology

Nature Communications, 2016, vol. 7, issue 1, 1-11

Abstract: Abstract The RNA-binding protein TRBP is a central component of the Dicer complex. Despite a decade of biochemical and structural studies, the essential functionality of TRBP in microRNA (miRNA) biogenesis remains unknown. Here we show that TRBP is an integral cofactor for time-efficient Dicer processing in RNA-crowded environments. We competed for Dicer processing of pre-miRNA with a large amount of cellular RNA species and found that Dicer-TRBP, but not Dicer alone, remains resilient. To apprehend the mechanism of this substrate selectivity, we use single-molecule fluorescence. The real-time observation reveals that TRBP acts as a gatekeeper, precluding Dicer from engaging with pre-miRNA-like substrates. TRBP acquires the selectivity using the PAZ domain of Dicer, whereas Dicer moderates the RNA-binding affinity of TRBP for fast turnover. This coordinated action between TRBP and Dicer accomplishes an efficient way of discarding pre-miRNA-like substrates.

Date: 2016
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DOI: 10.1038/ncomms13694

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