An EHBP-1-SID-3-DYN-1 axis promotes membranous tubule fission during endocytic recycling

PLoS Genetics
Jinghu GaoAnbing Shi

Abstract

The ACK family tyrosine kinase SID-3 is involved in the endocytic uptake of double-stranded RNA. Here we identified SID-3 as a previously unappreciated recycling regulator in the Caenorhabditis elegans intestine. The RAB-10 effector EHBP-1 is required for the endosomal localization of SID-3. Accordingly, animals with loss of SID-3 phenocopied the recycling defects observed in ehbp-1 and rab-10 single mutants. Moreover, we detected sequential protein interactions between EHBP-1, SID-3, NCK-1, and DYN-1. In the absence of SID-3, DYN-1 failed to localize at tubular recycling endosomes, and membrane tubules breaking away from endosomes were mostly absent, suggesting that SID-3 acts synergistically with the downstream DYN-1 to promote endosomal tubule fission. In agreement with these observations, overexpression of DYN-1 significantly increased recycling transport in SID-3-deficient cells. Finally, we noticed that loss of RAB-10 or EHBP-1 compromised feeding RNAi efficiency in multiple tissues, implicating basolateral recycling in the transport of RNA silencing signals. Taken together, our study demonstrated that in C. elegans intestinal epithelia, SID-3 acts downstream of EHBP-1 to direct fission of recycling endosomal tubules in c...Continue Reading

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Citations

Oct 13, 2020·RNA Biology·Fabian BraukmannEric Alexander Miska
Dec 19, 2020·The Journal of Biological Chemistry·Trey FarmerSteve Caplan
Dec 14, 2021·Frontiers in Cell and Developmental Biology·Shimin WangAnbing Shi

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Methods Mentioned

BETA
confocal microscopy
GTPase
Co-immunoprecipitation
pulldown
transgenic
PCR
in vitro transcription
electrophoresis
immunoprecipitation
fluorescence imaging

Software Mentioned

Elements AR
Fiji ( Image J )
Metamorph
GraphPad
Prism
NIS
JACoP ( Just Another Co - localization Plugin )

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