Modulation of lamellipodial structure and dynamics by NO-dependent phosphorylation of VASP Ser239
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Modulation of lamellipodial structure and dynamics by NO-dependent phosphorylation of VASP Ser239. / Lindsay, Susan L; Ramsey, Sara; Aitchison, Michael; Renné, Thomas; Evans, Thomas J.
In: J CELL SCI, Vol. 120, No. Pt 17, 01.09.2007, p. 3011-21.Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review
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TY - JOUR
T1 - Modulation of lamellipodial structure and dynamics by NO-dependent phosphorylation of VASP Ser239
AU - Lindsay, Susan L
AU - Ramsey, Sara
AU - Aitchison, Michael
AU - Renné, Thomas
AU - Evans, Thomas J
PY - 2007/9/1
Y1 - 2007/9/1
N2 - The initial step in directed cell movement is lamellipodial protrusion, an action driven by actin polymerization. Enabled/vasodilator-stimulated phosphoprotein (Ena/VASP) family proteins are key regulators of this actin polymerization and can control lamellipodial protrusion rate. Ena/VASP proteins are substrates for modification by cyclic-nucleotide-dependent protein kinases at a number of sites. Phosphorylation of Ser239 of VASP in vitro inhibits its anti-capping and filament-bundling activity but the effects of this modification on lamellipodial structure and function are unknown. To examine the functional effects of this modification in living cells, we studied VASP phosphorylation at Ser239 by nitric oxide (NO) stimulation of cGMP-dependent protein kinase. Using live cell imaging of primary cells transfected with GFP-VASP constructs, we found that NO produced rapid retraction of lamellipodia together with cell rounding that was dependent on guanylate cyclase and type II cGMP-dependent protein kinase. In cells expressing a mutant VASP (Ser239Ala) lacking the site preferentially phosphorylated by this kinase, NO had no effect. Phosphorylation of Ser239 of VASP results in loss of lamellipodial protrusions and cell rounding, and is a powerful means of controlling directed actin polymerization within lamellipodia.
AB - The initial step in directed cell movement is lamellipodial protrusion, an action driven by actin polymerization. Enabled/vasodilator-stimulated phosphoprotein (Ena/VASP) family proteins are key regulators of this actin polymerization and can control lamellipodial protrusion rate. Ena/VASP proteins are substrates for modification by cyclic-nucleotide-dependent protein kinases at a number of sites. Phosphorylation of Ser239 of VASP in vitro inhibits its anti-capping and filament-bundling activity but the effects of this modification on lamellipodial structure and function are unknown. To examine the functional effects of this modification in living cells, we studied VASP phosphorylation at Ser239 by nitric oxide (NO) stimulation of cGMP-dependent protein kinase. Using live cell imaging of primary cells transfected with GFP-VASP constructs, we found that NO produced rapid retraction of lamellipodia together with cell rounding that was dependent on guanylate cyclase and type II cGMP-dependent protein kinase. In cells expressing a mutant VASP (Ser239Ala) lacking the site preferentially phosphorylated by this kinase, NO had no effect. Phosphorylation of Ser239 of VASP results in loss of lamellipodial protrusions and cell rounding, and is a powerful means of controlling directed actin polymerization within lamellipodia.
KW - Actins
KW - Cell Adhesion Molecules
KW - Cell Shape
KW - Cells, Cultured
KW - Cyclic GMP-Dependent Protein Kinases
KW - Cytoskeleton
KW - Epithelial Cells
KW - Guanylate Cyclase
KW - Humans
KW - Kidney
KW - Microfilament Proteins
KW - Nitric Oxide
KW - Phosphoproteins
KW - Phosphorylation
KW - Pseudopodia
KW - Recombinant Fusion Proteins
KW - Serine
U2 - 10.1242/jcs.003061
DO - 10.1242/jcs.003061
M3 - SCORING: Journal article
C2 - 17684063
VL - 120
SP - 3011
EP - 3021
JO - J CELL SCI
JF - J CELL SCI
SN - 0021-9533
IS - Pt 17
ER -
