Activation of endogenous Cdc42 visualized in living cells.
Signaling proteins are tightly regulated spatially and temporally to perform multiple functions. For Cdc42 and other guanosine triphosphatases, the subcellular location of activation is a critical determinant of cell behavior. However, current approaches are limited in their ability to examine the dynamics of Cdc42 activity in living cells. We report the development of a biosensor capable of visualizing the changing activation of endogenous, unlabeled Cdc42 in living cells. With the use of a dye that reports protein interactions, the biosensor revealed localized activation in the trans-Golgi apparatus, microtubule-dependent Cdc42 activation at the cell periphery, and activation kinetics precisely coordinated with cell extension and retraction.
Nalbant
Perihan
P
Department of Pharmacology, University of North Carolina School of Medicine, Chapel Hill, NC 27599-7365, USA.
Hodgson
Louis
L
Kraynov
Vadim
V
Toutchkine
Alexei
A
Hahn
Klaus M
KM
D000199:Actins / Q000378:metabolism; D000465:Algorithms; D000818:Animals; D015374:Biosensing Techniques; D002448:Cell Adhesion; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism*; D016764:Cell Polarity; D022081:Cell Surface Extensions / Q000378:metabolism / Q000648:ultrastructure; D042783:Endothelial Cells / Q000378:metabolism / Q000648:ultrastructure; D005347:Fibroblasts; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000737:chemistry / Q000378:metabolism; D049452:Green Fluorescent Proteins; D006801:Humans; D008164:Luminescent Proteins; D051379:Mice; D008870:Microtubules / Q000378:metabolism; D018375:Neutrophil Activation; D009504:Neutrophils / Q000378:metabolism*; D011506:Proteins / Q000737:chemistry / Q000378:metabolism; D011554:Pseudopodia / Q000378:metabolism; D011744:Pyrimidinones / Q000378:metabolism; D012680:Sensitivity and Specificity; D051316:Wiskott-Aldrich Syndrome Protein; D020764:cdc42 GTP-Binding Protein / Q000378:metabolism*; D020741:rho GTP-Binding Proteins / Q000378:metabolism; D021601:trans-Golgi Network / Q000378:metabolism* / Q000648:ultrastructure
D000199:Actins; D005456:Fluorescent Dyes; D008164:Luminescent Proteins; D011506:Proteins; D011744:Pyrimidinones; C495767:WAS protein, human; C495768:Was protein, mouse; D051316:Wiskott-Aldrich Syndrome Protein; D049452:Green Fluorescent Proteins; C003954:merocyanine dye; C503256:RHOQ protein, human; C503260:Rhoq protein, mouse; D020764:cdc42 GTP-Binding Protein; D020741:rho GTP-Binding Proteins