title: A ZIP6-ZIP10 heteromer controls NCAM1 phosphorylation and integration into focal adhesion complexes during epithelial-to-mesenchymal transition.
abstract: The prion protein (PrP) evolved from the subbranch of ZIP metal ion transporters comprising ZIPs 5, 6 and 10, raising the prospect that the study of these ZIPs may reveal insights relevant for understanding the function of PrP. Building on data which suggested PrP and ZIP6 are critical during epithelial-to-mesenchymal transition (EMT), we investigated ZIP6 in an EMT paradigm using ZIP6 knockout cells, mass spectrometry and bioinformatic methods. Reminiscent of PrP, ZIP6 levels are five-fold upregulated during EMT and the protein forms a complex with NCAM1. ZIP6 also interacts with ZIP10 and the two ZIP transporters exhibit interdependency during their expression. ZIP6 contributes to the integration of NCAM1 in focal adhesion complexes but, unlike cells lacking PrP, ZIP6 deficiency does not abolish polysialylation of NCAM1. Instead, ZIP6 mediates phosphorylation of NCAM1 on a cluster of cytosolic acceptor sites. Substrate consensus motif features and in vitro phosphorylation data point toward GSK3 as the kinase responsible, and interface mapping experiments identified histidine-rich cytoplasmic loops within the ZIP6/ZIP10 heteromer as a novel scaffold for GSK3 binding. Our data suggests that PrP and ZIP6 inherited the ability to interact with NCAM1 from their common ZIP ancestors but have since diverged to control distinct posttranslational modifications of NCAM1.
authors.affiliation: Tanz Centre for Research in Neurodegenerative Diseases, University of Toronto, Ontario, Canada.|Department of Laboratory Medicine &Pathobiology, University of Toronto, Ontario, Canada.
authors.forename: Dylan
authors.initials: D
authors.lastname: Brethour
authors.affiliation: Tanz Centre for Research in Neurodegenerative Diseases, University of Toronto, Ontario, Canada.|Department of Laboratory Medicine &Pathobiology, University of Toronto, Ontario, Canada.
authors.forename: Mohadeseh
authors.initials: M
authors.lastname: Mehrabian
authors.affiliation: Tanz Centre for Research in Neurodegenerative Diseases, University of Toronto, Ontario, Canada.
authors.forename: Declan
authors.initials: D
authors.lastname: Williams
authors.affiliation: Tanz Centre for Research in Neurodegenerative Diseases, University of Toronto, Ontario, Canada.|Department of Laboratory Medicine &Pathobiology, University of Toronto, Ontario, Canada.
authors.forename: Xinzhu
authors.initials: X
authors.lastname: Wang
authors.affiliation: Tanz Centre for Research in Neurodegenerative Diseases, University of Toronto, Ontario, Canada.|Department of Laboratory Medicine &Pathobiology, University of Toronto, Ontario, Canada.
authors.forename: Farinaz
authors.initials: F
authors.lastname: Ghodrati
authors.affiliation: Tanz Centre for Research in Neurodegenerative Diseases, University of Toronto, Ontario, Canada.|Whitehead Institute for Biomedical Research, Cambridge, MA, USA.
authors.forename: Sepehr
authors.initials: S
authors.lastname: Ehsani
authors.affiliation: Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, Toronto, Ontario, Canada.
authors.forename: Elizabeth A
authors.initials: EA
authors.lastname: Rubie
authors.affiliation: Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, Toronto, Ontario, Canada.
authors.forename: James R
authors.initials: JR
authors.lastname: Woodgett
authors.affiliation: Tanz Centre for Research in Neurodegenerative Diseases, University of Toronto, Ontario, Canada.
authors.forename: Jean
authors.initials: J
authors.lastname: Sevalle
authors.affiliation: Tanz Centre for Research in Neurodegenerative Diseases, University of Toronto, Ontario, Canada.
authors.forename: Zhengrui
authors.initials: Z
authors.lastname: Xi
authors.affiliation: Tanz Centre for Research in Neurodegenerative Diseases, University of Toronto, Ontario, Canada.|Department of Neurology, University of Toronto, Ontario, Canada.
authors.forename: Ekaterina
authors.initials: E
authors.lastname: Rogaeva
authors.affiliation: Tanz Centre for Research in Neurodegenerative Diseases, University of Toronto, Ontario, Canada.|Department of Laboratory Medicine &Pathobiology, University of Toronto, Ontario, Canada.
authors.forename: Gerold
authors.initials: G
authors.lastname: Schmitt-Ulms
mesh_terms: D000199:Actins / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D019002:CD56 Antigen / Q000378:metabolism*; D027682:Cation Transport Proteins / Q000737:chemistry / Q000378:metabolism*; D003599:Cytoskeleton / Q000378:metabolism; D058750:Epithelial-Mesenchymal Transition; D022001:Focal Adhesions / Q000378:metabolism*; D038362:Glycogen Synthase Kinase 3 / Q000378:metabolism; D006639:Histidine / Q000378:metabolism; D006801:Humans; D016023:Integrins / Q000378:metabolism; D051379:Mice; D008954:Models, Biological; D019158:N-Acetylneuraminic Acid / Q000378:metabolism; D010766:Phosphorylation; D000072002:Prion Proteins / Q000378:metabolism; D011485:Protein Binding; D000072417:Protein Domains; D025941:Protein Interaction Mapping; D020033:Protein Isoforms / Q000737:chemistry / Q000378:metabolism; D017433:Protein Structure, Secondary
chemical_list: D000199:Actins; D019002:CD56 Antigen; D027682:Cation Transport Proteins; D016023:Integrins; C501063:Ncam1 protein, mouse; D000072002:Prion Proteins; D020033:Protein Isoforms; C454113:Slc39a6 protein, mouse; C557566:ZIP10 protein, mouse; D006639:Histidine; D038362:Glycogen Synthase Kinase 3; D019158:N-Acetylneuraminic Acid