Activation of endogenous Cdc42 visualized in living cells |
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| Authors: | Nalbant Perihan Hodgson Louis Kraynov Vadim Toutchkine Alexei Hahn Klaus M |
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| Institution: | Department of Pharmacology, University of North Carolina School of Medicine, Chapel Hill, NC 27599-7365, USA. |
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| Abstract: | 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. |
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