SIlva-Alvim, FAL, An, J, Alvim, JC et al. (6 more authors) (2018) Predominant Golgi-residency of the plant K/HDEL receptor is essential for its function in mediating ER retention. Plant Cell, 30 (9). pp. 2174-2196. ISSN 1040-4651
Abstract
Accumulation of soluble proteins in the endoplasmic reticulum (ER) of plants is mediated by a receptor termed ER RETENTION DEFECTIVE 2 (ERD2) or K/HDEL receptor. Using two gain-of-function assays and by complementing loss of function in Nicotiana benthamiana we discovered that compromising the lumenal N-terminus or the cytosolic C-terminus with fluorescent fusions abolishes its biological function and profoundly affects its subcellular localization. Based on the confirmed asymmetrical topology of ERD2 we engineered a new fluorescent ERD2 fusion protein that retains biological activity. Using this fusion, we show that ERD2 is exclusively detected at the Golgi apparatus, unlike non-functional C-terminal fusions which also label the ER. Moreover, ERD2 is confined to early Golgi compartments and does not show ligand-induced redistribution to the ER. We show that the cytosolic C-terminus of ERD2 plays a crucial role in its function. Two conserved Leucine residues that do not correspond to any known targeting motifs for ER-Golgi trafficking were shown to be essential for both ERD2 Golgi residency and its ability to mediate ER retention of soluble ligands. The results suggest that anterograde ER to Golgi transport of ERD2 is either extremely fast, well in excess of the bulk flow rate, or that ERD2 does not recycle in the way originally proposed.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | © 2018 The authors(s). Article free via Creative Commons CC-BY 4.0 license. (https://creativecommons.org/licenses/by/4.0/) |
Dates: |
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Institution: | The University of Leeds |
Academic Units: | The University of Leeds > Faculty of Biological Sciences (Leeds) > School of Biology (Leeds) |
Funding Information: | Funder Grant number BBSRC BB/D016223/1 Leverhulme Trust F/10 105/E |
Depositing User: | Symplectic Publications |
Date Deposited: | 29 Jun 2018 10:43 |
Last Modified: | 19 Oct 2018 14:24 |
Status: | Published |
Publisher: | American Society of Plant Biologists |
Identification Number: | 10.1105/tpc.18.00426 |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:132754 |