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Exo70E2 is essential for exocyst subunit recruitment and EXPO formation in both plants and animals

In contrast to a single copy of Exo70 in yeast and mammals, the Arabidopsis genome contains 23 paralogues of Exo70 (AtExo70). Using AtExo70E2 and its GFP fusion as probes, we recently identified a novel double-membrane organelle termed exocyst-positive organelle (EXPO) that mediates an unconventiona...

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Autores principales: Ding, Yu, Wang, Juan, Chun Lai, John Ho, Ling Chan, Vivian Hoi, Wang, Xiangfeng, Cai, Yi, Tan, Xiaoyun, Bao, Yiqun, Xia, Jun, Robinson, David G., Jiang, Liwen
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The American Society for Cell Biology 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3907280/
https://www.ncbi.nlm.nih.gov/pubmed/24307681
http://dx.doi.org/10.1091/mbc.E13-10-0586
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author Ding, Yu
Wang, Juan
Chun Lai, John Ho
Ling Chan, Vivian Hoi
Wang, Xiangfeng
Cai, Yi
Tan, Xiaoyun
Bao, Yiqun
Xia, Jun
Robinson, David G.
Jiang, Liwen
author_facet Ding, Yu
Wang, Juan
Chun Lai, John Ho
Ling Chan, Vivian Hoi
Wang, Xiangfeng
Cai, Yi
Tan, Xiaoyun
Bao, Yiqun
Xia, Jun
Robinson, David G.
Jiang, Liwen
author_sort Ding, Yu
collection PubMed
description In contrast to a single copy of Exo70 in yeast and mammals, the Arabidopsis genome contains 23 paralogues of Exo70 (AtExo70). Using AtExo70E2 and its GFP fusion as probes, we recently identified a novel double-membrane organelle termed exocyst-positive organelle (EXPO) that mediates an unconventional protein secretion in plant cells. Here we further demonstrate that AtExo70E2 is essential for exocyst subunit recruitment and for EXPO formation in both plants and animals. By performing transient expression in Arabidopsis protoplasts, we established that a number of exocyst subunits (especially the members of the Sec family) are unable to be recruited to EXPO in the absence of AtExo70E2. The paralogue AtExo70A1 is unable to substitute for AtExo70E2 in this regard. Fluorescence resonance energy transfer assay and bimolecular fluorescence complementation analyses confirm the interaction between AtExo70E2 and Sec6 and Sec10. AtExo70E2, but not its yeast counterpart, is also capable of inducing EXPO formation in an animal cell line (HEK293A cells). Electron microscopy confirms the presence of double-membraned, EXPO-like structures in HEK293A cells expressing AtExo70E2. Inversely, neither human nor yeast Exo70 homologues cause the formation of EXPO in Arabidopsis protoplasts. These results point to a specific and crucial role for AtExo70E2 in EXPO formation.
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spelling pubmed-39072802014-04-16 Exo70E2 is essential for exocyst subunit recruitment and EXPO formation in both plants and animals Ding, Yu Wang, Juan Chun Lai, John Ho Ling Chan, Vivian Hoi Wang, Xiangfeng Cai, Yi Tan, Xiaoyun Bao, Yiqun Xia, Jun Robinson, David G. Jiang, Liwen Mol Biol Cell Articles In contrast to a single copy of Exo70 in yeast and mammals, the Arabidopsis genome contains 23 paralogues of Exo70 (AtExo70). Using AtExo70E2 and its GFP fusion as probes, we recently identified a novel double-membrane organelle termed exocyst-positive organelle (EXPO) that mediates an unconventional protein secretion in plant cells. Here we further demonstrate that AtExo70E2 is essential for exocyst subunit recruitment and for EXPO formation in both plants and animals. By performing transient expression in Arabidopsis protoplasts, we established that a number of exocyst subunits (especially the members of the Sec family) are unable to be recruited to EXPO in the absence of AtExo70E2. The paralogue AtExo70A1 is unable to substitute for AtExo70E2 in this regard. Fluorescence resonance energy transfer assay and bimolecular fluorescence complementation analyses confirm the interaction between AtExo70E2 and Sec6 and Sec10. AtExo70E2, but not its yeast counterpart, is also capable of inducing EXPO formation in an animal cell line (HEK293A cells). Electron microscopy confirms the presence of double-membraned, EXPO-like structures in HEK293A cells expressing AtExo70E2. Inversely, neither human nor yeast Exo70 homologues cause the formation of EXPO in Arabidopsis protoplasts. These results point to a specific and crucial role for AtExo70E2 in EXPO formation. The American Society for Cell Biology 2014-02-01 /pmc/articles/PMC3907280/ /pubmed/24307681 http://dx.doi.org/10.1091/mbc.E13-10-0586 Text en © 2014 Ding 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 of Cell Biology.
spellingShingle Articles
Ding, Yu
Wang, Juan
Chun Lai, John Ho
Ling Chan, Vivian Hoi
Wang, Xiangfeng
Cai, Yi
Tan, Xiaoyun
Bao, Yiqun
Xia, Jun
Robinson, David G.
Jiang, Liwen
Exo70E2 is essential for exocyst subunit recruitment and EXPO formation in both plants and animals
title Exo70E2 is essential for exocyst subunit recruitment and EXPO formation in both plants and animals
title_full Exo70E2 is essential for exocyst subunit recruitment and EXPO formation in both plants and animals
title_fullStr Exo70E2 is essential for exocyst subunit recruitment and EXPO formation in both plants and animals
title_full_unstemmed Exo70E2 is essential for exocyst subunit recruitment and EXPO formation in both plants and animals
title_short Exo70E2 is essential for exocyst subunit recruitment and EXPO formation in both plants and animals
title_sort exo70e2 is essential for exocyst subunit recruitment and expo formation in both plants and animals
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3907280/
https://www.ncbi.nlm.nih.gov/pubmed/24307681
http://dx.doi.org/10.1091/mbc.E13-10-0586
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