mRNA structural dynamics shape Argonaute-target interactions

Journal article (2020)

Authors

Suzan Ruijtenberg University Medical Center Utrecht
Stijn Sonneveld University Medical Center Utrecht
T.J. Cui Kavli institute of nanoscience Delft, BN/Chirlmin Joo Lab - AS
Ive Logister University Medical Center Utrecht
Dion de Steenwinkel University Medical Center Utrecht
Yao Xiao The Scripps Research Institute, La Jolla
Ian J. MacRae The Scripps Research Institute, La Jolla
C. Joo BN/Chirlmin Joo Lab - AS , Kavli institute of nanoscience Delft
Marvin E. Tanenbaum University Medical Center Utrecht
Research Group
BN/Chirlmin Joo Lab (AS) (TU Delft)
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Published Date

2020

Language

English

Reuse Rights

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Faculty

Applied Sciences

Department

BN/Bionanoscience

Research Group

BN/Chirlmin Joo Lab

Abstract

Small interfering RNAs (siRNAs) promote RNA degradation in a variety of processes and have important clinical applications. siRNAs direct cleavage of target RNAs by guiding Argonaute2 (AGO2) to its target site. Target site accessibility is critical for AGO2-target interactions, but how target site accessibility is controlled in vivo is poorly understood. Here, we use live-cell single-molecule imaging in human cells to determine rate constants of the AGO2 cleavage cycle in vivo. We find that the rate-limiting step in mRNA cleavage frequently involves unmasking of target sites by translating ribosomes. Target site masking is caused by heterogeneous intramolecular RNA-RNA interactions, which can conceal target sites for many minutes in the absence of translation. Our results uncover how dynamic changes in mRNA structure shape AGO2-target recognition, provide estimates of mRNA folding and unfolding rates in vivo, and provide experimental evidence for the role of mRNA structural dynamics in control of mRNA-protein interactions.