title: EHBP1L1 coordinates Rab8 and Bin1 to regulate apical-directed transport in polarized epithelial cells.
abstract: The highly conserved Rab guanosine triphosphatase (GTPase) Rab8 plays a role in exocytosis toward the polarized plasma membrane in eukaryotic cells. In murine Rab8-deficient small intestine cells, apical proteins are missorted into lysosomes. In this study, we identified a novel Rab8-interacting protein complex containing an EH domain-binding protein 1-like 1 (EHBP1L1), Bin1/amphiphysin II, and dynamin. Biochemical analyses showed that EHBP1L1 directly bound to GTP-loaded Rab8 and Bin1. The spatial dependency of these complexes at the endocytic recycling compartment (ERC) was demonstrated through overexpression and knockdown experiments. EHBP1L1- or Bin1-depleted or dynamin-inhibited small intestine organoids significantly accumulated apical membrane proteins but not basolateral membrane proteins in lysosomes. Furthermore, in EHBP1L1-deficient mice, small intestine cells displayed truncated and sparse microvilli, suggesting that EHBP1L1 maintains the apical plasma membrane by regulating apical transport. In summary, our data demonstrate that EHBP1L1 links Rab8 and the Bin1-dynamin complex, which generates membrane curvature and excises the vesicle at the ERC for apical transport.
authors.affiliation: Department of Cell Biology, Graduate School of Medicine, Osaka University, Osaka 565-0871, Japan.
authors.forename: Atsuhiro
authors.initials: A
authors.lastname: Nakajo
authors.affiliation: Department of Cell Biology, Graduate School of Medicine, Osaka University, Osaka 565-0871, Japan shyoshimura@acb.med.osaka-u.ac.jp aharada@acb.med.osaka-u.ac.jp.
authors.forename: Shin-ichiro
authors.initials: S
authors.lastname: Yoshimura
authors.affiliation: Department of Cell Biology, Graduate School of Medicine, Osaka University, Osaka 565-0871, Japan.
authors.forename: Hiroko
authors.initials: H
authors.lastname: Togawa
authors.affiliation: Department of Cell Biology, Graduate School of Medicine, Osaka University, Osaka 565-0871, Japan.
authors.forename: Masataka
authors.initials: M
authors.lastname: Kunii
authors.affiliation: Department of Cell Biology, Graduate School of Medicine, Osaka University, Osaka 565-0871, Japan.
authors.forename: Tomohiko
authors.initials: T
authors.lastname: Iwano
authors.affiliation: Department of Cell Biology, Graduate School of Medicine, Osaka University, Osaka 565-0871, Japan.
authors.forename: Ayaka
authors.initials: A
authors.lastname: Izumi
authors.affiliation: Department of Cell Biology, Graduate School of Medicine, Osaka University, Osaka 565-0871, Japan.
authors.forename: Yuria
authors.initials: Y
authors.lastname: Noguchi
authors.affiliation: Department of Cell Biology, Graduate School of Medicine, Osaka University, Osaka 565-0871, Japan.
authors.forename: Ayako
authors.initials: A
authors.lastname: Watanabe
authors.affiliation: Department of Cell Biology, Graduate School of Medicine, Osaka University, Osaka 565-0871, Japan.
authors.forename: Ayako
authors.initials: A
authors.lastname: Goto
authors.affiliation: Department of Gastroenterology, Keio University School of Medicine, Tokyo 160-8582, Japan.
authors.forename: Toshiro
authors.initials: T
authors.lastname: Sato
authors.affiliation: Department of Cell Biology, Graduate School of Medicine, Osaka University, Osaka 565-0871, Japan shyoshimura@acb.med.osaka-u.ac.jp aharada@acb.med.osaka-u.ac.jp.
authors.forename: Akihiro
authors.initials: A
authors.lastname: Harada
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D001692:Biological Transport; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D016764:Cell Polarity; D034281:Dynamins / Q000378:metabolism; D004847:Epithelial Cells / Q000201:enzymology* / Q000648:ultrastructure; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D007413:Intestinal Mucosa / Q000201:enzymology* / Q000648:ultrastructure; D007421:Intestine, Small / Q000201:enzymology* / Q000648:ultrastructure; D008247:Lysosomes / Q000201:enzymology; D051379:Mice; D018345:Mice, Knockout; D008871:Microvilli / Q000201:enzymology; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism*; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D009940:Organoids; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D021381:Protein Transport; D034622:RNA Interference; D015398:Signal Transduction; D014162:Transfection; D022161:Transport Vesicles / Q000201:enzymology*; D025521:Tumor Suppressor Proteins / Q000235:genetics / Q000378:metabolism*; D020691:rab GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D048868:Adaptor Proteins, Signal Transducing; C101410:BIN1 protein, human; C109221:Bin1 protein, mouse; D002352:Carrier Proteins; C000608234:EHBP1L1 protein, human; C000608235:EHBP1L1 protein, mouse; D009419:Nerve Tissue Proteins; D009687:Nuclear Proteins; C521976:Rab8 protein, mouse; D025521:Tumor Suppressor Proteins; C082377:RAB8A protein, human; D020691:rab GTP-Binding Proteins; D034281:Dynamins
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