Differential VASP phosphorylation controls remodeling of the actin cytoskeleton
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Differential VASP phosphorylation controls remodeling of the actin cytoskeleton. / Benz, Peter M; Blume, Constanze; Seifert, Stefanie; Wilhelm, Sabine; Waschke, Jens; Schuh, Kai; Gertler, Frank; Münzel, Thomas; Renné, Thomas.
in: J CELL SCI, Jahrgang 122, Nr. Pt 21, 01.11.2009, S. 3954-65.Publikationen: SCORING: Beitrag in Fachzeitschrift/Zeitung › SCORING: Zeitschriftenaufsatz › Forschung › Begutachtung
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T1 - Differential VASP phosphorylation controls remodeling of the actin cytoskeleton
AU - Benz, Peter M
AU - Blume, Constanze
AU - Seifert, Stefanie
AU - Wilhelm, Sabine
AU - Waschke, Jens
AU - Schuh, Kai
AU - Gertler, Frank
AU - Münzel, Thomas
AU - Renné, Thomas
PY - 2009/11/1
Y1 - 2009/11/1
N2 - Proteins of the Enabled/vasodilator-stimulated phosphoprotein (Ena/VASP) family link signal transduction pathways to actin cytoskeleton dynamics. VASP is substrate of cAMP-dependent, cGMP-dependent and AMP-activated protein kinases that primarily phosphorylate the sites S157, S239 and T278, respectively. Here, we systematically analyzed functions of VASP phosphorylation patterns for actin assembly and subcellular targeting in vivo and compared the phosphorylation effects of Ena/VASP family members. Methods used were the reconstitution of VASP-null cells with ;locked' phosphomimetic VASP mutants, actin polymerization of VASP mutants in vitro and in living cells, site-specific kinase-mediated VASP phosphorylation, and analysis of the endogenous protein with phosphorylation-status-specific antibodies. Phosphorylation at S157 influenced VASP localization, but had a minor impact on F-actin assembly. Phosphorylation of the S157-equivalent site in the Ena/VASP family members Mena and EVL had no effect on the ratio of cellular F-actin to G-actin. By contrast, VASP phosphorylation at S239 (and the equivalent site in Mena) or T278 impaired VASP-driven actin filament formation. The data show that VASP functions are precisely regulated by differential phosphorylation and provide new insights into cytoskeletal control by serine/threonine kinase-dependent signaling pathways.
AB - Proteins of the Enabled/vasodilator-stimulated phosphoprotein (Ena/VASP) family link signal transduction pathways to actin cytoskeleton dynamics. VASP is substrate of cAMP-dependent, cGMP-dependent and AMP-activated protein kinases that primarily phosphorylate the sites S157, S239 and T278, respectively. Here, we systematically analyzed functions of VASP phosphorylation patterns for actin assembly and subcellular targeting in vivo and compared the phosphorylation effects of Ena/VASP family members. Methods used were the reconstitution of VASP-null cells with ;locked' phosphomimetic VASP mutants, actin polymerization of VASP mutants in vitro and in living cells, site-specific kinase-mediated VASP phosphorylation, and analysis of the endogenous protein with phosphorylation-status-specific antibodies. Phosphorylation at S157 influenced VASP localization, but had a minor impact on F-actin assembly. Phosphorylation of the S157-equivalent site in the Ena/VASP family members Mena and EVL had no effect on the ratio of cellular F-actin to G-actin. By contrast, VASP phosphorylation at S239 (and the equivalent site in Mena) or T278 impaired VASP-driven actin filament formation. The data show that VASP functions are precisely regulated by differential phosphorylation and provide new insights into cytoskeletal control by serine/threonine kinase-dependent signaling pathways.
KW - Actins
KW - Animals
KW - Cell Adhesion Molecules
KW - Cell Line
KW - Cytoskeleton
KW - Humans
KW - Mice
KW - Mice, Inbred C57BL
KW - Mice, Knockout
KW - Microfilament Proteins
KW - Phosphoproteins
KW - Phosphorylation
KW - Protein Transport
U2 - 10.1242/jcs.044537
DO - 10.1242/jcs.044537
M3 - SCORING: Journal article
C2 - 19825941
VL - 122
SP - 3954
EP - 3965
JO - J CELL SCI
JF - J CELL SCI
SN - 0021-9533
IS - Pt 21
ER -
