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PMID: 10953005 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Stimulation of fascin spikes by thrombospondin-1 is mediated by the GTPases Rac and Cdc42.

The Journal of cell biology ·Vol. 150 ·No. 4 ·2000-08-21 ·Pages 807-22

Adams JC, Schwartz MA

Abstract

Cell adhesion to extracellular matrix is an important physiological stimulus for organization of the actin-based cytoskeleton. Adhesion to the matrix glycoprotein thrombospondin-1 (TSP-1) triggers the sustained formation of F-actin microspikes that contain the actin-bundling protein fascin. These structures are also implicated in cell migration, which may be an important function of TSP-1 in tissue remodelling and wound repair. To further understand the function of fascin microspikes, we examined whether their assembly is regulated by Rho family GTPases. We report that expression of constitutively active mutants of Rac or Cdc42 triggered localization of fascin to lamellipodia, filopodia, and cell edges in fibroblasts or myoblasts. Biochemical assays demonstrated prolonged activation of Rac and Cdc42 in C2C12 cells adherent to TSP-1 and activation of the downstream kinase p21-activated kinase (PAK). Expression of dominant-negative Rac or Cdc42 in C2C12 myoblasts blocked spreading and formation of fascin spikes on TSP-1. Spreading and spike assembly were also blocked by pharmacological inhibition of F-actin turnover. Shear-loading of monospecific anti-fascin immunoglobulins, which block the binding of fascin to actin into cytoplasm, strongly inhibited spreading, actin cytoskeletal organization and migration on TSP-1 and also affected the motility of cells on fibronectin. We conclude that fascin is a critical component downstream of Rac and Cdc42 that is needed for actin cytoskeletal organization and cell migration responses to thrombospondin-1.

MeSH Terms
3T3 Cells Actins/metabolism Animals Bridged Bicyclo Compounds, Heterocyclic/pharmacology Carrier Proteins/metabolism Cell Adhesion/drug effects Cell Line Depsipeptides Fibronectins/physiology Mice Microfilament Proteins/metabolism Muscle, Skeletal/cytology,physiology Peptides, Cyclic/pharmacology Recombinant Proteins/metabolism Stress, Mechanical Thiazoles/pharmacology Thiazolidines Thrombospondin 1/physiology Transfection Vinculin/metabolism cdc42 GTP-Binding Protein/metabolism rac GTP-Binding Proteins/metabolism
Chemicals
Actins Bridged Bicyclo Compounds, Heterocyclic Carrier Proteins Depsipeptides Fibronectins Microfilament Proteins Peptides, Cyclic Recombinant Proteins Thiazoles Thiazolidines Thrombospondin 1 jasplakinolide Vinculin fascin cdc42 GTP-Binding Protein rac GTP-Binding Proteins latrunculin B
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Adams J C
MRC Laboratory for Molecular Cell Biology and Department of Biochemistry and Molecular Biology, University College London, London WC1E 6BT, United Kingdom. dmcbjca@ucl.ac.uk
Schwartz M A
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.150.4.807
Email
dmcbjca@ucl.ac.uk
Published
2000-08-21
Pages
807-22
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2175285
Subset
IM
Grants
NIGMS NIH HHS · R01 GM47214 · United States
Corrections
CommentIn
PMID: 10953018
External Links
PubMed source DOI: 10.1083/jcb.150.4.807 PMC Full Text View journal details
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