NDLI: Rho small GTPase regulates the stability of individual focal adhesions: a FRET-based visualization of GDP/GTP exchange on small GTPases

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Rho small GTPase regulates the stability of individual focal adhesions: a FRET-based visualization of GDP/GTP exchange on small GTPases

Content Provider	Semantic Scholar
Author	Miyauchi, Takayuki Yanagida, Toshio Sako, Yasushi
Copyright Year	2007
Abstract	RhoA and Rac1 are small GTPases primarily involved in cytoskeletal remodeling. Many biochemical studies have suggested that they are also key organizers of cell-substrate adhesion. Recently, fluorescence resonance energy transfer (FRET)-based indicators have been developed to visualize RhoA and Rac1 activity in living cells [Yoshizaki et al., J. Cell Biol. 162, 223 (2003); Pertz et al., Nature 440, 1069 (2006)]. These indicators use one of the interactions between RhoA (Rac1) and the RhoA (Rac1)-binding domain of their effector proteins. However, distribution of RhoA activity in single cells has not yet been observed with micrometer-scale resolution. Here, we employed an approach that detects GDP/GTP exchange on small GTPases by using FRET from YFP-fused small GTPases to a fluorescent analogue of GTP, BODIPY(TR)-GTP. This approach allowed us to visualize confined localization of active (GTP-bound forms of) RhoA and Rac1 in individual focal adhesions. Activated RhoA accumulated in immobile and long-lived focal adhesions but was not evident in unstable and temporary adhesions, while activated Rac1 was observed at every adhesion. Our results suggest that RhoA is the major regulator determining the stability of individual cell adhesion structures.
Starting Page	63
Ending Page	73
Page Count	11
File Format	PDF HTM / HTML
PubMed reference number	27857568v1
Volume Number	3
Journal	Biophysics
Language	English
Access Restriction	Open
Subject Keyword	Analog Cell Adhesion Cytoskeleton Fluorescence Resonance Energy Transfer Focal Adhesions Guanosine Diphosphate Guanosine Triphosphate Phosphohydrolases Imagery Micron Monomeric GTP-Binding Proteins Non-Small Cell Lung Carcinoma Tissue Adhesions Unstable Medical Device Problem VCL gene rhoA GTP-Binding Protein
Content Type	Text
Resource Type	Article

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