TY - JOUR
T1 - Argonaute bypasses cellular obstacles without hindrance during target search
AU - Cui, Tao Ju
AU - Klein, Misha
AU - Hegge, Jorrit W.
AU - Chandradoss, Stanley D.
AU - van der Oost, John
AU - Depken, Martin
AU - Joo, Chirlmin
N1 - Funding Information:
We thank Ian MacRae for critical reading and Malwina Szczepaniak, Margreet Docter, Jelle van der Does, Dimitri de Roos, Anna Haagsma and Jan Wignand for technical support. C.J. was supported by Vidi (864.14.002) of the Netherlands Organization for Scientific research and an ERC Consolidator Grant (819299) of the European Research Council. M.K. and M.D. were supported by the Netherlands Organization for Scientific Research, as part of the Frontiers in Nanoscience program. M.D. acknowledges financial support from a TU Delft startup grant. J.v.d.O. was financially supported by two grants from the Netherlands Organization of Scientific Research (NWO; ECHO grant 711.013.002 and NWO-TOP grant 714.015.001).
Publisher Copyright:
© 2019, The Author(s).
PY - 2019/12/1
Y1 - 2019/12/1
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=85072699819&partnerID=8YFLogxK
U2 - 10.1038/s41467-019-12415-y
DO - 10.1038/s41467-019-12415-y
M3 - Article
C2 - 31558728
AN - SCOPUS:85072699819
SN - 2041-1723
VL - 10
JO - Nature Communications
JF - Nature Communications
IS - 1
M1 - 4390
ER -