title: ARHGEF10 directs the localization of Rab8 to Rab6-positive executive vesicles.
abstract: The function of ARHGEF10, a known guanine nucleotide exchange factor (GEF) for RhoA with proposed roles in various diseases, is poorly understood. To understand the precise function of this protein, we raised a monoclonal antibody against ARHGEF10 and determined its localization in HeLa cells. ARHGEF10 was found to localize to vesicles containing Rab6 (of which there are three isoforms, Rab6a, Rab6b and Rab6c), Rab8 (of which there are two isoforms, Rab8a and Rab8b), and/or the secretion marker neuropeptide Y (NPY)-Venus in a Rab6-dependent manner. These vesicles were known to originate from the Golgi and contain secreted or membrane proteins. Ectopic expression of an N-terminal-truncated ARHGEF10 mutant led to the generation of large vesicle-like structures containing both Rab6 and Rab8. Additionally, small interfering (si)RNA-mediated knockdown of ARHGEF10 impaired the localization of Rab8 to these exocytotic vesicles. Furthermore, the invasiveness of MDA-MB231 cells was markedly decreased by knockdown of ARHGEF10, as well as of Rab8. From these results, we propose that ARHGEF10 acts in exocytosis and tumor invasion in a Rab8-dependent manner.
authors.affiliation: Group of Neurobiology, Division of Health Sciences, Graduate School of Medicine, Osaka University, Osaka 565-0871, Japan sshibata@sahs.med.osaka-u.ac.jp inagaki@sahs.med.osaka-u.ac.jp.
authors.forename: Satoshi
authors.initials: S
authors.lastname: Shibata
authors.affiliation: Group of Neurobiology, Division of Health Sciences, Graduate School of Medicine, Osaka University, Osaka 565-0871, Japan.
authors.forename: Tsubasa
authors.initials: T
authors.lastname: Kawanai
authors.affiliation: Group of Neurobiology, Division of Health Sciences, Graduate School of Medicine, Osaka University, Osaka 565-0871, Japan.
authors.forename: Takayuki
authors.initials: T
authors.lastname: Hara
authors.affiliation: Group of Neurobiology, Division of Health Sciences, Graduate School of Medicine, Osaka University, Osaka 565-0871, Japan.
authors.forename: Asuka
authors.initials: A
authors.lastname: Yamamoto
authors.affiliation: Group of Neurobiology, Division of Health Sciences, Graduate School of Medicine, Osaka University, Osaka 565-0871, Japan.
authors.forename: Taro
authors.initials: T
authors.lastname: Chaya
authors.affiliation: Group of Neurobiology, Division of Health Sciences, Graduate School of Medicine, Osaka University, Osaka 565-0871, Japan.
authors.forename: Yasunori
authors.initials: Y
authors.lastname: Tokuhara
authors.affiliation: Group of Neurobiology, Division of Health Sciences, Graduate School of Medicine, Osaka University, Osaka 565-0871, Japan.
authors.forename: Chinami
authors.initials: C
authors.lastname: Tsuji
authors.affiliation: Group of Neurobiology, Division of Health Sciences, Graduate School of Medicine, Osaka University, Osaka 565-0871, Japan.
authors.forename: Manabu
authors.initials: M
authors.lastname: Sakai
authors.affiliation: Department of Bioengineering, Graduate School of Engineering, Osaka City University, Osaka 558-8585, Japan.
authors.forename: Taro
authors.initials: T
authors.lastname: Tachibana
authors.affiliation: Group of Neurobiology, Division of Health Sciences, Graduate School of Medicine, Osaka University, Osaka 565-0871, Japan sshibata@sahs.med.osaka-u.ac.jp inagaki@sahs.med.osaka-u.ac.jp.
authors.forename: Shinobu
authors.identifier: 0000-0002-2785-1214
authors.initials: S
authors.lastname: Inagaki
mesh_terms: D008841:Actin Cytoskeleton / Q000378:metabolism; D000911:Antibodies, Monoclonal / Q000378:metabolism; D000918:Antibody Specificity / Q000276:immunology; D045744:Cell Line, Tumor; D005089:Exocytosis; D055785:Gene Knockdown Techniques; D006801:Humans; D050505:Mutant Proteins / Q000378:metabolism; D009361:Neoplasm Invasiveness; D009478:Neuropeptide Y / Q000378:metabolism; D058105:Polymerization; D000072417:Protein Domains; D021381:Protein Transport; D064067:Rho Guanine Nucleotide Exchange Factors / Q000378:metabolism*; D022142:Secretory Vesicles / Q000378:metabolism*; D020691:rab GTP-Binding Proteins / Q000378:metabolism*
chemical_list: C577232:ARHGEF10 protein, human; D000911:Antibodies, Monoclonal; D050505:Mutant Proteins; D009478:Neuropeptide Y; C094135:Rab6 protein; D064067:Rho Guanine Nucleotide Exchange Factors; C082377:RAB8A protein, human; D020691:rab GTP-Binding Proteins
keywords: ARHGEF10; Membrane trafficking; RabGTPase; RhoGEF