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Argonaute bypasses cellular obstacles without hindrance during target search

Tao Ju Cui, Misha Klein, Jorrit Hegge, Stanley D Chandradoss, John van der Oost, Martin Depken, Chirlmin Joo
doi: https://doi.org/10.1101/535575
Tao Ju Cui
Delft University of Technology;
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Misha Klein
Delft University of Technology;
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Jorrit Hegge
Wageningen University;
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Stanley D Chandradoss
Oxford NanoImaging Ltd
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John van der Oost
Wageningen University;
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Martin Depken
Delft University of Technology;
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Chirlmin Joo
Delft University of Technology;
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  • For correspondence: c.joo@tudelft.nl
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Abstract

Argonaute (Ago) proteins are key players in gene regulation in eukaryotes and host defense in prokaryotes. For specific interference, Ago relies on base pairing between small nucleic acid guides and complementary target sequences. To efficiently scan nucleic acid chains for potential targets, Ago must bypass both secondary structures in mRNA and single stranded DNA as well as protein barriers. Through single-molecule FRET, we reveal that lateral diffusion is mediated mainly through protein-nucleic acid interactions, rather than interactions between the guide and targeted strand. This allows Ago to scan for targets with high efficiency but without maintaining tight contact with the DNA backbone. Real-time observations show that Ago glides short distances over secondary structures while using intersegmental jumps to reduce scanning redundancy and bypass protein barriers. Our single-molecule method in combination with kinetic analysis may serve as a novel platform to study the effect of sequence on search kinetics for other nucleic acid-guided proteins.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-ND 4.0 International license.
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Posted January 30, 2019.
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Argonaute bypasses cellular obstacles without hindrance during target search
Tao Ju Cui, Misha Klein, Jorrit Hegge, Stanley D Chandradoss, John van der Oost, Martin Depken, Chirlmin Joo
bioRxiv 535575; doi: https://doi.org/10.1101/535575
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Argonaute bypasses cellular obstacles without hindrance during target search
Tao Ju Cui, Misha Klein, Jorrit Hegge, Stanley D Chandradoss, John van der Oost, Martin Depken, Chirlmin Joo
bioRxiv 535575; doi: https://doi.org/10.1101/535575

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