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Conserved function of the lysine-based KXD/E motif in Golgi retention for endomembrane proteins among different organisms

We recently identified a new COPI-interacting KXD/E motif in the C-terminal cytosolic tail (CT) of Arabidopsis endomembrane protein 12 (AtEMP12) as being a crucial Golgi retention mechanism for AtEMP12. This KXD/E motif is conserved in CTs of all EMPs found in plants, yeast, and humans and is also p...

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Autores principales: Woo, Cheuk Hang, Gao, Caiji, Yu, Ping, Tu, Linna, Meng, Zhaoyue, Banfield, David K., Yao, Xiaoqiang, Jiang, Liwen
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The American Society for Cell Biology 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4642860/
https://www.ncbi.nlm.nih.gov/pubmed/26378254
http://dx.doi.org/10.1091/mbc.E15-06-0361
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author Woo, Cheuk Hang
Gao, Caiji
Yu, Ping
Tu, Linna
Meng, Zhaoyue
Banfield, David K.
Yao, Xiaoqiang
Jiang, Liwen
author_facet Woo, Cheuk Hang
Gao, Caiji
Yu, Ping
Tu, Linna
Meng, Zhaoyue
Banfield, David K.
Yao, Xiaoqiang
Jiang, Liwen
author_sort Woo, Cheuk Hang
collection PubMed
description We recently identified a new COPI-interacting KXD/E motif in the C-terminal cytosolic tail (CT) of Arabidopsis endomembrane protein 12 (AtEMP12) as being a crucial Golgi retention mechanism for AtEMP12. This KXD/E motif is conserved in CTs of all EMPs found in plants, yeast, and humans and is also present in hundreds of other membrane proteins. Here, by cloning selective EMP isoforms from plants, yeast, and mammals, we study the localizations of EMPs in different expression systems, since there are contradictory reports on the localizations of EMPs. We show that the N-terminal and C-terminal GFP-tagged EMP fusions are localized to Golgi and post-Golgi compartments, respectively, in plant, yeast, and mammalian cells. In vitro pull-down assay further proves the interaction of the KXD/E motif with COPI coatomer in yeast. COPI loss of function in yeast and plants causes mislocalization of EMPs or KXD/E motif–containing proteins to vacuole. Ultrastructural studies further show that RNA interference (RNAi) knockdown of coatomer expression in transgenic Arabidopsis plants causes severe morphological changes in the Golgi. Taken together, our results demonstrate that N-terminal GFP fusions reflect the real localization of EMPs, and KXD/E is a conserved motif in COPI interaction and Golgi retention in eukaryotes.
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spelling pubmed-46428602016-01-30 Conserved function of the lysine-based KXD/E motif in Golgi retention for endomembrane proteins among different organisms Woo, Cheuk Hang Gao, Caiji Yu, Ping Tu, Linna Meng, Zhaoyue Banfield, David K. Yao, Xiaoqiang Jiang, Liwen Mol Biol Cell Articles We recently identified a new COPI-interacting KXD/E motif in the C-terminal cytosolic tail (CT) of Arabidopsis endomembrane protein 12 (AtEMP12) as being a crucial Golgi retention mechanism for AtEMP12. This KXD/E motif is conserved in CTs of all EMPs found in plants, yeast, and humans and is also present in hundreds of other membrane proteins. Here, by cloning selective EMP isoforms from plants, yeast, and mammals, we study the localizations of EMPs in different expression systems, since there are contradictory reports on the localizations of EMPs. We show that the N-terminal and C-terminal GFP-tagged EMP fusions are localized to Golgi and post-Golgi compartments, respectively, in plant, yeast, and mammalian cells. In vitro pull-down assay further proves the interaction of the KXD/E motif with COPI coatomer in yeast. COPI loss of function in yeast and plants causes mislocalization of EMPs or KXD/E motif–containing proteins to vacuole. Ultrastructural studies further show that RNA interference (RNAi) knockdown of coatomer expression in transgenic Arabidopsis plants causes severe morphological changes in the Golgi. Taken together, our results demonstrate that N-terminal GFP fusions reflect the real localization of EMPs, and KXD/E is a conserved motif in COPI interaction and Golgi retention in eukaryotes. The American Society for Cell Biology 2015-11-15 /pmc/articles/PMC4642860/ /pubmed/26378254 http://dx.doi.org/10.1091/mbc.E15-06-0361 Text en © 2015 Woo, Gao, et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0). “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society for Cell Biology.
spellingShingle Articles
Woo, Cheuk Hang
Gao, Caiji
Yu, Ping
Tu, Linna
Meng, Zhaoyue
Banfield, David K.
Yao, Xiaoqiang
Jiang, Liwen
Conserved function of the lysine-based KXD/E motif in Golgi retention for endomembrane proteins among different organisms
title Conserved function of the lysine-based KXD/E motif in Golgi retention for endomembrane proteins among different organisms
title_full Conserved function of the lysine-based KXD/E motif in Golgi retention for endomembrane proteins among different organisms
title_fullStr Conserved function of the lysine-based KXD/E motif in Golgi retention for endomembrane proteins among different organisms
title_full_unstemmed Conserved function of the lysine-based KXD/E motif in Golgi retention for endomembrane proteins among different organisms
title_short Conserved function of the lysine-based KXD/E motif in Golgi retention for endomembrane proteins among different organisms
title_sort conserved function of the lysine-based kxd/e motif in golgi retention for endomembrane proteins among different organisms
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4642860/
https://www.ncbi.nlm.nih.gov/pubmed/26378254
http://dx.doi.org/10.1091/mbc.E15-06-0361
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