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PMID: 15361624 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Activation of endogenous Cdc42 visualized in living cells.

Science (New York, N.Y.) ·Vol. 305 ·No. 5690 ·2004-09-10 ·Pages 1615-9

Nalbant P, Hodgson L, Kraynov V, Toutchkine A, Hahn KM

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.

MeSH Terms

Actins/metabolism Algorithms Animals Biosensing Techniques Cell Adhesion Cell Line Cell Membrane/metabolism Cell Polarity Cell Surface Extensions/metabolism,ultrastructure Endothelial Cells/metabolism,ultrastructure Fibroblasts Fluorescence Fluorescent Dyes/chemistry,metabolism Green Fluorescent Proteins Humans Luminescent Proteins Mice Microtubules/metabolism Neutrophil Activation Neutrophils/metabolism Proteins/chemistry,metabolism Pseudopodia/metabolism Pyrimidinones/metabolism Sensitivity and Specificity Wiskott-Aldrich Syndrome Protein cdc42 GTP-Binding Protein/metabolism rho GTP-Binding Proteins/metabolism trans-Golgi Network/metabolism,ultrastructure

Chemicals

Actins Fluorescent Dyes Luminescent Proteins Proteins Pyrimidinones WAS protein, human Was protein, mouse Wiskott-Aldrich Syndrome Protein Green Fluorescent Proteins merocyanine dye RHOQ protein, human Rhoq protein, mouse cdc42 GTP-Binding Protein rho GTP-Binding Proteins

Authors & Affiliations

5 authors, click to expand affiliations / ORCID

Nalbant Perihan

Department of Pharmacology, University of North Carolina School of Medicine, Chapel Hill, NC 27599-7365, USA.

Hodgson Louis

Kraynov Vadim

Toutchkine Alexei

Hahn Klaus M

Article Info

Journal

Science (New York, N.Y.)

Abbr.

Science

ISSN

1095-9203

DOI

10.1126/science.1100367

Published

2004-09-10

Pages

1615-9

Language

English

Region

United States

NLM ID

0404511

Subset

Grants

NIGMS NIH HHS · R01 GM057464 · United States

NIGMS NIH HHS · GM57464 · United States

NIGMS NIH HHS · GM64346 · United States

External Links

PubMed source DOI: 10.1126/science.1100367 View journal details

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Activation of endogenous Cdc42 visualized in living cells.

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