Argonaute bypasses cellular obstacles without hindrance during target search

Journal article (2019)

Authors

Thijs Cui Kavli institute of nanoscience Delft
M. Klein BN/Chirlmin Joo Lab - AS
Jorrit W. Hegge Wageningen University & Research
S.D. Chandradoss BN/Chirlmin Joo Lab - AS , Kavli institute of nanoscience Delft
John van der Oost Wageningen University & Research
S.M. Depken Kavli institute of nanoscience Delft, BN/Martin Depken Lab - AS
C. Joo Kavli institute of nanoscience Delft, BN/Chirlmin Joo Lab - AS
Research Group
BN/Martin Depken Lab (AS) (TU Delft)
Copyright: © 2019 Thijs Cui, M. Klein, Jorrit W. Hegge, S.D. Chandradoss, John van der Oost, S.M. Depken, C. Joo
DOI
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https://doi.org/10.1038/s41467-019-12415-y
More Info
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Published Date

2019

Language

English

Faculty

Applied Sciences

Department

BN/Bionanoscience

Research Group

BN/Martin Depken Lab

Abstract

Argonaute (Ago) proteins are key players in both gene regulation (eukaryotes) and host defense (prokaryotes). Acting on single-stranded nucleic-acid substrates, Ago relies on base pairing between a small nucleic-acid guide and its complementary target sequences for specificity. To efficiently scan nucleic-acid chains for targets, Ago diffuses laterally along the substrate and must bypass secondary structures as well as protein barriers. Using single-molecule FRET in conjunction with kinetic modelling, we reveal that target scanning is mediated through loose protein-nucleic acid interactions, allowing Ago to slide short distances over secondary structures, as well as to bypass protein barriers via intersegmental transfer. Our combined single-molecule experiment and kinetic modelling approach may serve as a platform to dissect search processes and study the effect of sequence on search kinetics for other nucleic acid-guided proteins.