title: Exo70E2 is essential for exocyst subunit recruitment and EXPO formation in both plants and animals.
abstract: 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.
authors.affiliation: School of Life Sciences, Centre for Cell and Developmental Biology and State Key Laboratory of Agrobiotechnology, Chinese University of Hong Kong, Shatin, New Territories, Hong Kong, China Division of Life Science, Division of Biomedical Engineering and State Key Laboratory of Molecular Neuroscience, Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong, China College of Life Sciences, Nanjing Agricultural University, Nanjing 210095, China Department of Plant Cell Biology, Centre for Organismal Studies, University of Heidelberg, D-69120 Heidelberg, Germany Shenzhen Research Institute, Chinese University of Hong Kong, Shenzhen 518057, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Ding
authors.forename: Juan
authors.initials: J
authors.lastname: Wang
authors.forename: John Ho
authors.initials: JH
authors.lastname: Chun Lai
authors.forename: Vivian Hoi
authors.initials: VH
authors.lastname: Ling Chan
authors.forename: Xiangfeng
authors.initials: X
authors.lastname: Wang
authors.forename: Yi
authors.initials: Y
authors.lastname: Cai
authors.forename: Xiaoyun
authors.initials: X
authors.lastname: Tan
authors.forename: Yiqun
authors.initials: Y
authors.lastname: Bao
authors.forename: Jun
authors.initials: J
authors.lastname: Xia
authors.forename: David G
authors.initials: DG
authors.lastname: Robinson
authors.forename: Liwen
authors.initials: L
authors.lastname: Jiang
mesh_terms: D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000235:genetics / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D005089:Exocytosis / Q000235:genetics; D031541:Fluorescence Resonance Energy Transfer; D015870:Gene Expression; D057809:HEK293 Cells; D006801:Humans; D008969:Molecular Sequence Data; D059828:Plant Cells; D011401:Promoter Regions, Genetic; D016415:Sequence Alignment; D015398:Signal Transduction / Q000235:genetics; D033921:Vesicular Transport Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism*
chemical_list: D029681:Arabidopsis Proteins; C547754:EXOC7 protein, human; C000592430:Exo70E2 protein, Arabidopsis; D033921:Vesicular Transport Proteins