title: ARHGAP22 localizes at endosomes and regulates actin cytoskeleton.
abstract: Rho small GTPases control cell morphology and motility through the rearrangement of actin cytoskeleton. We have previously shown that FilGAP, a Rac-specific GAP, binds to the actin-cross-linking protein Filamin A (FLNa) and suppresses Rac-dependent lamellae formation and cell spreading. ARHGAP22 is a member of FilGAP family, and implicated in the regulation of tumor cell motility. However, little is known concerning the cellular localization and mechanism of regulation at the molecular level. Whereas FilGAP binds to FLNa and localizes to lamellae, we found that ARHGAP22 did not bind to FLNa. Forced expression of ARHGAP22 induced enlarged vesicular structures containing the endocytic markers EEA1, Rab5, and Rab11. Moreover, endogenous ARHGAP22 is co-localized with EEA1- and Rab11-positive endosomes but not with trans-Golgi marker TNG46. When constitutively activated Rac Q61L mutant was expressed, ARHGAP22 is co-localized with Rac Q61L at membrane ruffles, suggesting that ARHGAP22 is translocated from endosomes to membrane ruffles to inactivate Rac. Forced expression of ARHGAP22 suppressed lamellae formation and cell spreading. Conversely, knockdown of endogenous ARHGAP22 stimulated cell spreading. Thus, our findings suggest that ARHGAP22 controls cell morphology by inactivating Rac but its localization is not mediated by its interaction with FLNa.
authors.affiliation: Division of Cell Biology, Department of Biosciences, School of Science, Kitasato University, Kanagawa, Japan.
authors.forename: Mamiko
authors.initials: M
authors.lastname: Mori
authors.affiliation: Division of Cell Biology, Department of Biosciences, School of Science, Kitasato University, Kanagawa, Japan.
authors.forename: Koji
authors.initials: K
authors.lastname: Saito
authors.affiliation: Division of Cell Biology, Department of Biosciences, School of Science, Kitasato University, Kanagawa, Japan.
authors.forename: Yasutaka
authors.initials: Y
authors.lastname: Ohta
mesh_terms: D008841:Actin Cytoskeleton / Q000378:metabolism*; D000818:Animals; D001665:Binding Sites; D015153:Blotting, Western; D002465:Cell Movement; D002478:Cells, Cultured; D011992:Endosomes / Q000378:metabolism*; D064448:Filamins / Q000378:metabolism*; D020690:GTPase-Activating Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D047468:Immunoprecipitation; D051379:Mice; D018482:Muscle, Skeletal / Q000166:cytology / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000235:genetics; D033921:Vesicular Transport Proteins / Q000378:metabolism; D020691:rab GTP-Binding Proteins / Q000378:metabolism; D020696:rab5 GTP-Binding Proteins / Q000378:metabolism; D020744:rac GTP-Binding Proteins / Q000378:metabolism*; D000099193:rab11 GTP-Binding Proteins
chemical_list: D064448:Filamins; D020690:GTPase-Activating Proteins; D034741:RNA, Small Interfering; D033921:Vesicular Transport Proteins; D020691:rab GTP-Binding Proteins; D020696:rab5 GTP-Binding Proteins; D020744:rac GTP-Binding Proteins; D000099193:rab11 GTP-Binding Proteins; C578338:ARHGAP22 protein, human; C093846:early endosome antigen 1; C427755:RAB5C protein, human