Single-molecule analysis of processive double-stranded RNA cleavage by Drosophila Dicer-2

Masahiro Naganuma, Hisashi Tadakuma () and Yukihide Tomari ()
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Masahiro Naganuma: The University of Tokyo
Hisashi Tadakuma: The University of Tokyo
Yukihide Tomari: The University of Tokyo

Nature Communications, 2021, vol. 12, issue 1, 1-9

Abstract: Abstract Drosophila Dicer-2 (Dcr-2) produces small interfering RNAs from long double-stranded RNAs (dsRNAs), playing an essential role in antiviral RNA interference. The dicing reaction by Dcr-2 is enhanced by Loquacious-PD (Loqs-PD), a dsRNA-binding protein that partners with Dcr-2. Previous biochemical analyses have proposed that Dcr-2 uses two distinct—processive or distributive—modes of cleavage by distinguishing the terminal structures of dsRNAs and that Loqs-PD alters the terminal dependence of Dcr-2. However, the direct evidence for this model is lacking, as the dynamic movement of Dcr-2 along dsRNAs has not been traced. Here, by utilizing single-molecule imaging, we show that the terminal structures of long dsRNAs and the presence or absence of Loqs-PD do not essentially change Dcr-2’s cleavage mode between processive and distributive, but rather simply affect the probability for Dcr-2 to undergo the cleavage reaction. Our results provide a refined model for how the dicing reaction by Dcr-2 is regulated.

Date: 2021
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DOI: 10.1038/s41467-021-24555-1

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