Integration of two RAB5 groups during endosomal transport in plants

ELife
Emi ItoTakashi Ueda

Abstract

RAB5 is a key regulator of endosomal functions in eukaryotic cells. Plants possess two different RAB5 groups, canonical and plant-unique types, which act via unknown counteracting mechanisms. Here, we identified an effector molecule of the plant-unique RAB5 in Arabidopsis thaliana, ARA6, which we designated PLANT-UNIQUE RAB5 EFFECTOR 2 (PUF2). Preferential colocalization with canonical RAB5 on endosomes and genetic interaction analysis indicated that PUF2 coordinates vacuolar transport with canonical RAB5, although PUF2 was identified as an effector of ARA6. Competitive binding of PUF2 with GTP-bound ARA6 and GDP-bound canonical RAB5, together interacting with the shared activating factor VPS9a, showed that ARA6 negatively regulates canonical RAB5-mediated vacuolar transport by titrating PUF2 and VPS9a. These results suggest a unique and unprecedented function for a RAB effector involving the integration of two RAB groups to orchestrate endosomal trafficking in plant cells.

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Citations

Jul 3, 2019·The Plant Journal : for Cell and Molecular Biology·Monika KaldeFarhah F Assaad
Apr 24, 2020·Proceedings of the National Academy of Sciences of the United States of America·María Otilia DelgadilloEnrique Rojo

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Methods Mentioned

BETA
GTPase
GTPases
two-hybrid
co-immunoprecipitation
transgenic
immunoprecipitation
pull-down
column chromatography

Software Mentioned

Fiji
ImageJ

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