title: Surface expression of the Anoctamin-1 (ANO1) channel is suppressed by protein-protein interactions with β-COP.
abstract: Anoctamin-1 (ANO1) is a Ca(2+)-activated chloride channel (CaCC) that plays important physiological roles in normal and cancerous tissues. However, the plasma membrane trafficking mechanisms of ANO1 remain poorly characterized. In yeast two-hybrid screening experiments, we observed direct interactions of ANO1 with β-COP, which is a subunit of Coat Protein Complex I (COPI). This interaction was then confirmed using several in vitro and in vivo binding assays. Moreover, the cotransfection of β-COP with ANO1 into HEK293T cells led to decreased the surface expression and the channel activity of ANO1. Accordingly, endogenous ANO1 was associated with β-COP in U251 glioblastoma cells, and silencing of β-COP enhanced surface expression and whole-cell currents of ANO1 in these cells. Taken together, these data suggest that β-COP negatively regulates ANO1 surface expression.
authors.affiliation: School of Biosystem and Biomedical Science, College of Health Science, Korea University, Seoul 02841, Republic of Korea; Center for Functional Connectomics, Korea Institute of Science and Technology (KIST), Seoul 02792, Republic of Korea.
authors.forename: Young-Sun
authors.initials: YS
authors.lastname: Lee
authors.affiliation: School of Biosystem and Biomedical Science, College of Health Science, Korea University, Seoul 02841, Republic of Korea.
authors.forename: Yeonju
authors.initials: Y
authors.lastname: Bae
authors.affiliation: School of Biosystem and Biomedical Science, College of Health Science, Korea University, Seoul 02841, Republic of Korea.
authors.forename: Nammi
authors.initials: N
authors.lastname: Park
authors.affiliation: School of Biosystem and Biomedical Science, College of Health Science, Korea University, Seoul 02841, Republic of Korea.
authors.forename: Jae Cheal
authors.initials: JC
authors.lastname: Yoo
authors.affiliation: School of Biosystem and Biomedical Science, College of Health Science, Korea University, Seoul 02841, Republic of Korea.
authors.forename: Chang-Hoon
authors.initials: CH
authors.lastname: Cho
authors.affiliation: School of Biosystem and Biomedical Science, College of Health Science, Korea University, Seoul 02841, Republic of Korea.
authors.forename: Kanghyun
authors.initials: K
authors.lastname: Ryoo
authors.affiliation: Center for Functional Connectomics, Korea Institute of Science and Technology (KIST), Seoul 02792, Republic of Korea; Neuroscience Program, University of Science and Technology (UST), Daejeon 34113, Republic of Korea. Electronic address: emhwang@kist.re.kr.
authors.forename: Eun Mi
authors.initials: EM
authors.lastname: Hwang
authors.affiliation: School of Biosystem and Biomedical Science, College of Health Science, Korea University, Seoul 02841, Republic of Korea. Electronic address: jaeyong68@korea.ac.kr.
authors.forename: Jae-Yong
authors.initials: JY
authors.lastname: Park
mesh_terms: D000075369:Anoctamin-1; D001692:Biological Transport; D001932:Brain Neoplasms / Q000378:metabolism; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism; D018118:Chloride Channels / Q000378:metabolism*; D020756:Coatomer Protein / Q000032:analysis / Q000378:metabolism*; D005909:Glioblastoma / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D009363:Neoplasm Proteins / Q000378:metabolism*; D060066:Protein Interaction Maps
chemical_list: C476213:ANO1 protein, human; D000075369:Anoctamin-1; C000619076:COPB1 protein, human; D018118:Chloride Channels; D020756:Coatomer Protein; D009363:Neoplasm Proteins
keywords: ANO1; Protein–protein interactions; Surface expression; U251 glioblastoma cells; Yeast two-hybrid screening; β-COP