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  • ROOT HAIR DEFECTIVE3 Is a Receptor for Selective Autophagy of the Endoplasmic Reticulum in Arabidopsis.

ROOT HAIR DEFECTIVE3 Is a Receptor for Selective Autophagy of the Endoplasmic Reticulum in Arabidopsis.

Frontiers in plant science (2022-03-15)
Jiaqi Sun, Weina Wang, Huanquan Zheng
ABSTRACT

ROOT HAIR DEFECTIVE3 (RHD3) is a plant member of atlastin GTPases, which belong to an evolutionally conserved family of proteins that mediate the homotypic fusion of the endoplasmic reticulum (ER). An atlastin in mammalian cells has recently been shown to act as an ER-phagy receptor for selective autophagy of the ER (ER-phagy) during nutrient starvation. Although RHD3 has been indicated to play a role in ER stress response, it is not very clear how RHD3 is involved in the process. In this study, we showed that the rhd3 mutant is hyposensitive to ER as well as salt stress. We employed an YFP-tagged ER membrane marker YFP-TMC to monitor the efficiency of ER-phagy microscopically and biochemically. We found that rhd3 is defective in ER-phagy under ER stress. Furthermore, there is an increased association of YFP-RHD3 with ATG8e-marked autophagosomes. YFP-RHD3 is also visible with ATG8e in the vacuole, and there is a breakdown of YFP-RHD3 under ER stress. RHD3 has two putative ATG8 interaction motifs (AIM1-2). We revealed that RHD3 but not RHD3(ΔAIM1) physically interacts with ATG8, a core autophagosomal component that interacts with various receptor proteins to recruit cargos for degradation by selective autophagy. Furthermore, their interaction is enhanced under ER stress. We thus propose that RHD3 acts as an ER-phagy receptor under ER stress to promote ER-phagy in Arabidopsis.

MATERIALS
Product Number
Brand
Product Description

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Anti-GFP, N-terminal antibody produced in rabbit, ~1.0 mg/mL, affinity isolated antibody, buffered aqueous solution
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Protease Inhibitor Cocktail, for plant cell and tissue extracts, DMSO solution
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Anti-α-Tubulin antibody, Mouse monoclonal, clone B-5-1-2, purified from hybridoma cell culture