1. Argonaute bypasses cellular obstacles without hindrance during target search
- Author
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Cui, Thijs (author), Klein, M. (author), Hegge, Jorrit W. (author), Chandradoss, S.D. (author), van der Oost, John (author), Depken, S.M. (author), Joo, C. (author), Cui, Thijs (author), Klein, M. (author), Hegge, Jorrit W. (author), Chandradoss, S.D. (author), van der Oost, John (author), Depken, S.M. (author), and Joo, C. (author)
- 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., BN/Chirlmin Joo Lab, BN/Martin Depken Lab
- Published
- 2019
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