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Characterization of DCL 4 missense alleles provides insights into its ability to process distinct classes of ds RNA substrates
Author(s) -
Montavon Thomas,
Kwon Yerim,
Zimmermann Aude,
Hammann Philippe,
Vincent Timothée,
Cognat Valérie,
Bergdoll Marc,
Michel Fabrice,
Dunoyer Patrice
Publication year - 2018
Publication title -
the plant journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.058
H-Index - 269
eISSN - 1365-313X
pISSN - 0960-7412
DOI - 10.1111/tpj.13941
Subject(s) - missense mutation , allele , rna , chemistry , biology , computational biology , genetics , gene , mutation
Summary In the model plant Arabidopsis thaliana , four Dicer‐like proteins ( DCL 1–4) mediate the production of various classes of small RNA s ( sRNA s). Among these four proteins, DCL 4 is by far the most versatile RN ase III ‐like enzyme, and previously identified dcl4 missense alleles were shown to uncouple the production of the various classes of DCL 4‐dependent sRNA s. Yet little is known about the molecular mechanism behind this uncoupling. Here, by studying the subcellular localization, interactome and binding to the sRNA precursors of three distinct dcl4 missense alleles, we simultaneously highlight the absolute requirement of a specific residue in the helicase domain for the efficient production of all DCL 4‐dependent sRNA s, and identify, within the PAZ domain, an important determinant of DCL 4 versatility that is mandatory for the efficient processing of intramolecular fold‐back double‐stranded RNA (ds RNA ) precursors, but that is dispensable for the production of small interfering RNA s (si RNA s) from RDR ‐dependent ds RNA susbtrates. This study not only provides insights into the DCL 4 mode of action, but also delineates interesting tools to further study the complexity of RNA silencing pathways in plants, and possibly other organisms.