Mechanical constraint imposed on plasma membrane through transverse phospholipid imbalance induces reversible actin polymerization via phosphoinositide 3-kinase activation.
Platelets were used to explore the effect of membrane curvature induced by phospholipid excess on cell shape and on organization of the actin cytoskeleton. We showed that the addition of short chain analogues of phospholipids to the outer leaflet of plasma membrane of resting platelets immediately induced a shape change with long filopodia formation containing newly polymerized actin. Cells recovered rapidly their discoid shape and their initial F-actin content only with the phosphatidylserine analogue, which was transported to the inner leaflet by aminophospholipid translocase. Filopodia formation and actin polymerization were inhibited in platelets pre-incubated with cytochalasin D. Both wortmannin and LY294002, two unrelated inhibitors of phosphoinositide 3-kinase, considerably reduced actin polymerization and filopodia formation. Phospholipid imbalance was accompanied by a reversible translocation of phosphoinositide 3-kinase from cytoplasm to plasma membrane. In agreement with a role for PI 3-kinase, when phospholipids were added to platelets, PtdIns(3,4)P2 increased two-fold and Akt protein was partly phosphorylated. A similar shape change was also observed in nocodazole-treated L929 fibroblasts which were incubated with the similar phospholipid analogues. In those nucleated cells, where the microtubule cytoskeleton was disrupted, a major actin-dependent membrane extension was induced by addition of short chain phospholipids that required the functional integrity of PI 3-kinase. We conclude that any physical constraint acting on plasma membrane and resulting on local changes in membrane curvature is sufficient to initiate transient actin polymerization via phosphoinositide 3-kinase activation.
Bettache
Nadir
N
CNRS-UMR 5539, Université Montpellier 2, Place Eugène Bataillon, 34095 Montpellier Cedex 05, France. bettache@univ-montp2.fr
Baisamy
Laurent
L
Baghdiguian
Stephen
S
Payrastre
Bernard
B
Mangeat
Paul
P
Bienvenue
Alain
A
D008841:Actin Cytoskeleton / Q000378:metabolism*; D000730:Androstadienes / Q000494:pharmacology; D001792:Blood Platelets / Q000502:physiology*; D002462:Cell Membrane / Q000502:physiology*; D002867:Chromones / Q000494:pharmacology; D004789:Enzyme Activation / Q000502:physiology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005347:Fibroblasts; D006801:Humans; D009025:Morpholines / Q000494:pharmacology; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism*; D000081082:Phosphoinositide-3 Kinase Inhibitors; D010743:Phospholipids / Q000378:metabolism*; D010766:Phosphorylation; D011108:Polymers / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases; D011518:Proto-Oncogene Proteins / Q000378:metabolism; D051057:Proto-Oncogene Proteins c-akt; D011554:Pseudopodia / Q000502:physiology; D000077191:Wortmannin
D000730:Androstadienes; D002867:Chromones; D004791:Enzyme Inhibitors; D009025:Morpholines; D019869:Phosphatidylinositol 3-Kinases; D000081082:Phosphoinositide-3 Kinase Inhibitors; D010743:Phospholipids; D011108:Polymers; D017346:Protein Serine-Threonine Kinases; D011518:Proto-Oncogene Proteins; D051057:Proto-Oncogene Proteins c-akt; D000077191:Wortmannin; C085911:2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one; C494918:AKT1 protein, human