title: In vitro and in vivo characterization of the Rab11-GAP activity of Drosophila Evi5.
abstract: Small GTPases of the Rab family are master regulators of vesicular trafficking. As such, they control the spatial distribution of various proteins, including proteins involved in cell signaling and the regulation of cell polarity. Misregulation of Rab proteins is associated with a large array of diseases. Surprisingly, the target of some key regulators of Rab proteins, including many GTPase-activating protein (GAP) is still unknown. Identifying the target of a specific GAP requires the combination of both in vitro and in vivo experiments to avoid any misinterpretation. Here is described the methodology we used to characterize the Rab11-GAP activity of Drosophila Evi5. We first focus on the in vitro Rab11 effector pull-down assay we developed and then we detail the in vivo characterization of Rab11 activity during Drosophila border cell migration.
authors: Institute for Research in Immunology and Cancer, University of Montréal, Montréal, QC, Canada, H3C 3J7.
authors: Carl
authors: C
authors: Laflamme
authors: Gregory
authors: G
authors: Emery
mesh_terms: D000818:Animals; D002460:Cell Line; D029721:Drosophila Proteins / Q000378:metabolism*; D004331:Drosophila melanogaster; D005260:Female; D020690:GTPase-Activating Proteins / Q000378:metabolism*; D057054:Molecular Imaging; D010053:Ovary / Q000378:metabolism; D020691:rab GTP-Binding Proteins / Q000378:metabolism*; D000099193:rab11 GTP-Binding Proteins
chemical_list: D029721:Drosophila Proteins; D020690:GTPase-Activating Proteins; D020691:rab GTP-Binding Proteins; C000605622:Evi5 protein, Drosophila; C559123:Rab11 protein, Drosophila; D000099193:rab11 GTP-Binding Proteins