title: Cadherin-related neuronal receptor 1 (CNR1) has cell adhesion activity with beta1 integrin mediated through the RGD site of CNR1.
abstract: Cadherin-related neuronal receptor (CNR) proteins are a diverse set of synaptic protocadherins, but little is known about its adhesive properties. We found that overexpressed CNR1 protein localized on the cell surface of HEK293T cells and increased the calcium-dependent cell aggregation potential. However, we could not detect the strong homophilic binding activity of CNR1 EC-Fc fusion protein in vitro. Parental HEK293T cells adhered to Arg-Gly-Asp (RGD) motif of EC1 domain of CNR1-Fc fusion protein. The fusion protein that the Asp73 of EC1 point-mutated to Glu (RGE-Fc) lost the adhesive activity. The adhesion activity of HEK293T cells to CNR1 EC-Fc fusion protein was completely blocked by inhibitors of integrins, including RGDS peptide and anti-beta1 integrin antibodies. The increased cell-aggregative property of CNR1 transfectants was also blocked by RGDS peptides. At cell-cell junctions of the CNR1 transfectants, co-localization between CNR1 and HEK293T endogenous beta1 integrin was observed. Furthermore, the spatiotemporal expression patterns of CNR and beta1 integrin nearly overlapped in the molecular layer of the developing mouse cerebellum in the main stage of synaptogenesis. These results indicate that CNR1 has a heterophilic, calcium-dependent cell adhesion activity with the beta1 integrin subfamily, and raise the possibility of CNR-beta1 integrin association in synaptogenesis.
authors.affiliation: KOKORO Biology Group, Laboratories for Integrated Biology, Graduate School of Frontier Biosciences, Osaka University, 1-3 Yamadaoka, Suita 565-0871, Japan. tmuto@brain.riken.go.jp
authors.forename: Tetsuji
authors.initials: T
authors.lastname: Mutoh
authors.forename: Shun
authors.initials: S
authors.lastname: Hamada
authors.forename: Kouji
authors.initials: K
authors.lastname: Senzaki
authors.forename: Yoji
authors.initials: Y
authors.lastname: Murata
authors.forename: Takeshi
authors.initials: T
authors.lastname: Yagi
mesh_terms: D000818:Animals; D000906:Antibodies / Q000494:pharmacology; D001665:Binding Sites / Q000502:physiology; D016466:CHO Cells; D019556:COS Cells; D020013:Calcium Signaling / Q000502:physiology; D002448:Cell Adhesion / Q000235:genetics; D015816:Cell Adhesion Molecules, Neuronal / Q000235:genetics / Q000378:metabolism*; D002454:Cell Differentiation / Q000235:genetics*; D002462:Cell Membrane / Q000235:genetics / Q000378:metabolism*; D002525:Cerebellar Cortex / Q000254:growth & development / Q000378:metabolism; D006224:Cricetinae; D004285:Dogs; D020439:Growth Cones / Q000378:metabolism*; D006801:Humans; D019012:Integrin beta1 / Q000187:drug effects / Q000378:metabolism*; D007365:Intercellular Junctions / Q000235:genetics / Q000378:metabolism; D051379:Mice; D009154:Mutation / Q000235:genetics; D009479:Neuropeptides / Q000235:genetics / Q000378:metabolism*; D010455:Peptides / Q000494:pharmacology; D017434:Protein Structure, Tertiary / Q000235:genetics; D000091363:Protocadherins; D011956:Receptors, Cell Surface / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D013569:Synapses / Q000235:genetics / Q000378:metabolism*
chemical_list: D000906:Antibodies; D015816:Cell Adhesion Molecules, Neuronal; D019012:Integrin beta1; D009479:Neuropeptides; C484820:PCDHA4 protein, human; C113143:Pcdha4 protein, mouse; D010455:Peptides; D000091363:Protocadherins; D011956:Receptors, Cell Surface; D011993:Recombinant Fusion Proteins