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A dual role for Rac1 GTPases in the regulation of cell motility

Filić, Vedrana; Marinović, Maja; Faix, Jan; Weber, Igor (2012) A dual role for Rac1 GTPases in the regulation of cell motility. Journal of Cell Science, 125 (2). pp. 387-398. ISSN 0021-9533

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Rac proteins are the only canonical Rho family GTPases in Dictyostelium, where they act as key regulators of the actin cytoskeleton. In order to monitor the dynamics of activated Rac1 in Dictyostelium cells, a fluorescent probe was developed that specifically binds to GTP-bound form of Rac1. The probe is based on the GTPase- binding domain (GBD) from PAK1 kinase, and was selected on the basis of yeast two-hybrid, GST pull-down and fluorescence resonance energy transfer assays. PAK1_GBD localizes to leading edges of migrating cells and to endocytotic cups. As in vertebrates, activated Rac1 therefore appears to control de novo actin polymerization at protruding regions of the cell. Additionally, we found that the IQGAP-related protein DGAP1, which sequesters active Rac1 into a quaternary complex with actin-binding proteins cortexillin I and II, localizes to the trailing regions of migrating cells. Notably, PAK1_GBD and DGAP1, which both bind to Rac1-GTP, display mutually exclusive localizations in cell migration, phagocytosis and cytokinesis, and opposite dynamics of recruitment to the cell cortex upon stimulation with chemoattractants. Moreover, cortical localization of PAK1_GBD depends on integrity of the actin cytoskeleton, whereas cortical localization of DGAP1 does not. Taken together, these results imply that Rac1 GTPases play a dual role in regulation of cell motility and polarity in Dictyostelium.

Item Type: Article
Uncontrolled Keywords: Rac1; cell motility; cell polarization; FRET; actin cytoskeleton; Rho GTPases; dictyostelium
Subjects: NATURAL SCIENCES > Mathematics
Divisions: Division of Molecular Biology
Project titleProject leaderProject codeProject type
Unity through Knowledge FundUNSPECIFIED1A 10/07UNSPECIFIED
Regulacija dinamike citoskeleta u kretanju i diobi stanica-Igor Weber098-0982913-2858MZOS
Deutsche Akademische AustauschdienstUNSPECIFIEDDAAD-D/07/00065UNSPECIFIED
Deutsche ForschungsgemeinschaftUNSPECIFIEDDFG FA 330/4-2UNSPECIFIED
Depositing User: Igor Weber
Date Deposited: 13 Mar 2017 11:43
DOI: 10.1242/jcs.089680

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