Interaction of cortactin and Arp2/3 complex is required for sphingosine-1-phosphate-induced endothelial cell remodeling.
Sphingosine-1-phosphate (S1P) induces capillary formation of endothelial cells on Matrigel in accompany with actin assembly and accumulation of cortactin and Arp2/3 complex at the cell-leading edge. Suppression of cortactin expression with a cortactin antisense oligo significantly impaired S1P-induced capillary formation, migration of endothelial cells, and actin assembly at the cell periphery. Overexpression of wild-type cortactin tagged by green fluorescent protein (GFP) increased the S1P-induced tube formation and cell motility, whereas the cells overexpressing the mutant formed poorly capillary network and became less motile in response to S1P. Analysis of distribution in Triton X-100 insoluble fractions demonstrated that the cortactin mutant inhibited the association of wild-type cortactin and Arp2/3 complex with the actin-enriched complex. Furthermore, actin polymerization at and distribution of Arp2/3 complex as well as endogenous cortactin into the cell-leading edge mediated by S1P was disturbed. These data suggest that the interaction between cortactin and Arp2/3 complex plays an important role in S1P-mediated remodeling of endothelial cells.
Li
Yansong
Y
Department of Experimental Pathology, Jerome H. Holland Laboratory for the Biomedical Sciences, American Red Cross, Rockville, MD 20855, USA.
Uruno
Takehito
T
Haudenschild
Christian
C
Dudek
Steven M
SM
Garcia
Joe G N
JG
Zhan
Xi
X
D051377:Actin-Related Protein 2; D051378:Actin-Related Protein 3; D000199:Actins / Q000096:biosynthesis; D002196:Capillaries / Q000166:cytology / Q000254:growth & development / Q000378:metabolism; D002465:Cell Movement / Q000235:genetics; D051356:Cortactin; D003598:Cytoskeletal Proteins / Q000378:metabolism*; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D049452:Green Fluorescent Proteins; D006801:Humans; D008164:Luminescent Proteins; D008246:Lysophospholipids / Q000378:metabolism* / Q000494:pharmacology; D046911:Macromolecular Substances; D008840:Microfilament Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D009154:Mutation / Q000235:genetics; D018919:Neovascularization, Physiologic / Q000235:genetics*; D020319:Oligodeoxyribonucleotides, Antisense / Q000494:pharmacology; D011108:Polymers / Q000378:metabolism; D013110:Sphingosine / Q000031:analogs & derivatives / Q000378:metabolism* / Q000494:pharmacology; D015854:Up-Regulation / Q000235:genetics
C495926:ACTR2 protein, human; C495928:ACTR3 protein, human; D051377:Actin-Related Protein 2; D051378:Actin-Related Protein 3; D000199:Actins; C495868:CTTN protein, human; D051356:Cortactin; D003598:Cytoskeletal Proteins; D008164:Luminescent Proteins; D008246:Lysophospholipids; D046911:Macromolecular Substances; D008840:Microfilament Proteins; D020319:Oligodeoxyribonucleotides, Antisense; D011108:Polymers; D049452:Green Fluorescent Proteins; C060506:sphingosine 1-phosphate; D013110:Sphingosine