Regulation of podosome dynamics by WASp phosphorylation: Implication in matrix degradation and chemotaxis in macrophages

Athanassios Dovas, Jean Claude Gevrey, Alberto Grossi, Haein Park, Wassim Abou-Kheir, Dianne Cox

Research output: Contribution to journalArticle

74 Citations (Scopus)

Abstract

Podosomes, adhesion structures capable of matrix degradation, have been linked with the ability of cells to perform chemotaxis and invade tissues. Wiskott-Aldrich Syndrome protein (WASp), an effector of the RhoGTPase Cdc42 and a Src family kinase substrate, regulates macrophage podosome formation. In this study, we demonstrate that WASp is active in podosomes by using TIRF-FRET microscopy. Pharmacological and RNA interference approaches suggested that continuous WASp activity is required for podosome formation and function. Rescue experiments using point mutations demonstrate an absolute requirement for Cdc42 binding to WASp in podosome formation. Although tyrosine phosphorylation was not absolutely required for podosome formation, phosphorylation did regulate the rate of podosome nucleation and actin filament stability. Importantly, WASp tyrosine phosphorylation does not alter WASp activation, instead phosphorylation appears to be important for the restriction of WASp activity to podosomes. In addition, the matrix-degrading ability of cells requires WASp phosphorylation. Chemotactic responses to CSF-1 were also attenuated in the absence of endogenous WASp, which could not be rescued with either tyrosine mutation. These results suggest a more complex role for tyrosine phosphorylation than simply in the regulation of WASp activity, and suggest a link between podosome dynamics and macrophage migration.

Original languageEnglish (US)
Pages (from-to)3873-3882
Number of pages10
JournalJournal of Cell Science
Volume122
Issue number21
DOIs
StatePublished - 2009

Fingerprint

Wiskott-Aldrich Syndrome Protein
Chemotaxis
Macrophages
Phosphorylation
Tyrosine
Aptitude
Podosomes
Macrophage Colony-Stimulating Factor
src-Family Kinases
RNA Interference
Actin Cytoskeleton
Point Mutation
Microscopy

Keywords

  • Phosphorylation
  • Podosomes
  • WASp

ASJC Scopus subject areas

  • Cell Biology

Cite this

Regulation of podosome dynamics by WASp phosphorylation : Implication in matrix degradation and chemotaxis in macrophages. / Dovas, Athanassios; Gevrey, Jean Claude; Grossi, Alberto; Park, Haein; Abou-Kheir, Wassim; Cox, Dianne.

In: Journal of Cell Science, Vol. 122, No. 21, 2009, p. 3873-3882.

Research output: Contribution to journalArticle

Dovas, Athanassios ; Gevrey, Jean Claude ; Grossi, Alberto ; Park, Haein ; Abou-Kheir, Wassim ; Cox, Dianne. / Regulation of podosome dynamics by WASp phosphorylation : Implication in matrix degradation and chemotaxis in macrophages. In: Journal of Cell Science. 2009 ; Vol. 122, No. 21. pp. 3873-3882.
@article{cfabcb4bd66840b79fc38c66f185347d,
title = "Regulation of podosome dynamics by WASp phosphorylation: Implication in matrix degradation and chemotaxis in macrophages",
abstract = "Podosomes, adhesion structures capable of matrix degradation, have been linked with the ability of cells to perform chemotaxis and invade tissues. Wiskott-Aldrich Syndrome protein (WASp), an effector of the RhoGTPase Cdc42 and a Src family kinase substrate, regulates macrophage podosome formation. In this study, we demonstrate that WASp is active in podosomes by using TIRF-FRET microscopy. Pharmacological and RNA interference approaches suggested that continuous WASp activity is required for podosome formation and function. Rescue experiments using point mutations demonstrate an absolute requirement for Cdc42 binding to WASp in podosome formation. Although tyrosine phosphorylation was not absolutely required for podosome formation, phosphorylation did regulate the rate of podosome nucleation and actin filament stability. Importantly, WASp tyrosine phosphorylation does not alter WASp activation, instead phosphorylation appears to be important for the restriction of WASp activity to podosomes. In addition, the matrix-degrading ability of cells requires WASp phosphorylation. Chemotactic responses to CSF-1 were also attenuated in the absence of endogenous WASp, which could not be rescued with either tyrosine mutation. These results suggest a more complex role for tyrosine phosphorylation than simply in the regulation of WASp activity, and suggest a link between podosome dynamics and macrophage migration.",
keywords = "Phosphorylation, Podosomes, WASp",
author = "Athanassios Dovas and Gevrey, {Jean Claude} and Alberto Grossi and Haein Park and Wassim Abou-Kheir and Dianne Cox",
year = "2009",
doi = "10.1242/jcs.051755",
language = "English (US)",
volume = "122",
pages = "3873--3882",
journal = "Journal of Cell Science",
issn = "0021-9533",
publisher = "Company of Biologists Ltd",
number = "21",

}

TY - JOUR

T1 - Regulation of podosome dynamics by WASp phosphorylation

T2 - Implication in matrix degradation and chemotaxis in macrophages

AU - Dovas, Athanassios

AU - Gevrey, Jean Claude

AU - Grossi, Alberto

AU - Park, Haein

AU - Abou-Kheir, Wassim

AU - Cox, Dianne

PY - 2009

Y1 - 2009

N2 - Podosomes, adhesion structures capable of matrix degradation, have been linked with the ability of cells to perform chemotaxis and invade tissues. Wiskott-Aldrich Syndrome protein (WASp), an effector of the RhoGTPase Cdc42 and a Src family kinase substrate, regulates macrophage podosome formation. In this study, we demonstrate that WASp is active in podosomes by using TIRF-FRET microscopy. Pharmacological and RNA interference approaches suggested that continuous WASp activity is required for podosome formation and function. Rescue experiments using point mutations demonstrate an absolute requirement for Cdc42 binding to WASp in podosome formation. Although tyrosine phosphorylation was not absolutely required for podosome formation, phosphorylation did regulate the rate of podosome nucleation and actin filament stability. Importantly, WASp tyrosine phosphorylation does not alter WASp activation, instead phosphorylation appears to be important for the restriction of WASp activity to podosomes. In addition, the matrix-degrading ability of cells requires WASp phosphorylation. Chemotactic responses to CSF-1 were also attenuated in the absence of endogenous WASp, which could not be rescued with either tyrosine mutation. These results suggest a more complex role for tyrosine phosphorylation than simply in the regulation of WASp activity, and suggest a link between podosome dynamics and macrophage migration.

AB - Podosomes, adhesion structures capable of matrix degradation, have been linked with the ability of cells to perform chemotaxis and invade tissues. Wiskott-Aldrich Syndrome protein (WASp), an effector of the RhoGTPase Cdc42 and a Src family kinase substrate, regulates macrophage podosome formation. In this study, we demonstrate that WASp is active in podosomes by using TIRF-FRET microscopy. Pharmacological and RNA interference approaches suggested that continuous WASp activity is required for podosome formation and function. Rescue experiments using point mutations demonstrate an absolute requirement for Cdc42 binding to WASp in podosome formation. Although tyrosine phosphorylation was not absolutely required for podosome formation, phosphorylation did regulate the rate of podosome nucleation and actin filament stability. Importantly, WASp tyrosine phosphorylation does not alter WASp activation, instead phosphorylation appears to be important for the restriction of WASp activity to podosomes. In addition, the matrix-degrading ability of cells requires WASp phosphorylation. Chemotactic responses to CSF-1 were also attenuated in the absence of endogenous WASp, which could not be rescued with either tyrosine mutation. These results suggest a more complex role for tyrosine phosphorylation than simply in the regulation of WASp activity, and suggest a link between podosome dynamics and macrophage migration.

KW - Phosphorylation

KW - Podosomes

KW - WASp

UR - http://www.scopus.com/inward/record.url?scp=70450236976&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=70450236976&partnerID=8YFLogxK

U2 - 10.1242/jcs.051755

DO - 10.1242/jcs.051755

M3 - Article

C2 - 19808890

AN - SCOPUS:70450236976

VL - 122

SP - 3873

EP - 3882

JO - Journal of Cell Science

JF - Journal of Cell Science

SN - 0021-9533

IS - 21

ER -