Localization of urokinase type plasminogen activator to focal adhesions requires ligation of vitronectin integrin receptors.
Previous studies have shown that the adhesion protein, vitronectin, directs the localization of urokinase-type plasminogen activator (uPA) to areas of cell-substrate adhesion, where uPA is thought to regulate cell migration as well as pericellular proteolysis. In the present study, HT-1080 cell lines expressing either wild-type vitronectin or vitronectin containing a single amino-acid substitution in the integrin binding domain were used to assess whether ligation of the alphavbeta5 integrin was required for uPA localization to focal adhesions. The synthesis of wild-type vitronectin by HT-1080 cells adherent to either collagen or fibronectin resulted in the redistribution of both the alphavbeta5 integrin as well as uPA to focal adhesion structures. In contrast, cells synthesizing mutant vitronectin, containing the amino-acid substitution in the integrin binding domain, were unable to direct the redistribution of either alphavbeta5 or uPA to focal adhesions. Recombinant forms of wild-type and mutant vitronectin were prepared in a baculovirus system and compared for their ability to direct the redistribution of vitronectin integrin receptors as well as uPA on human skin fibroblasts. In the absence of vitronectin, fibroblast cells adherent to fibronectin assemble focal adhesions which contain the beta1 integrin but do not contain uPA. Addition of recombinant wild-type, but not mutant, vitronectin to fibroblasts adherent to fibronectin resulted in the redistribution of alphavbeta3, alphavbeta5, and uPA into focal adhesions. However, when cells were plated directly onto antibodies directed against either the alphavbeta3 or alphavbeta5 integrins, uPA was not localized on the cell surface. These data indicate that ligation of vitronectin integrin receptors is necessary but not sufficient for the localization of uPA to areas of cell matrix adhesion, and suggest that vitronectin may promote cell migration by recruiting vitronectin integrin receptors and components of the plasminogen activator system to areas of cell matrix contact.
Wilcox-Adelman
S A
SA
Center for Cell Biology and Cancer Research, Albany Medical College, New York 12208, USA.
Wilkins-Port
C E
CE
McKeown-Longo
P J
PJ
D005109:Extracellular Matrix / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D005354:Fibrosarcoma; D022001:Focal Adhesions / Q000737:chemistry* / Q000201:enzymology*; D015870:Gene Expression / Q000502:physiology; D006801:Humans; D016023:Integrins / Q000032:analysis / Q000378:metabolism; D008026:Ligation; D009842:Oligopeptides / Q000378:metabolism; D019030:Receptors, Vitronectin / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D012867:Skin / Q000166:cytology; D014162:Transfection; D014407:Tumor Cells, Cultured; D014568:Urokinase-Type Plasminogen Activator / Q000032:analysis* / Q000378:metabolism; D019096:Vitronectin / Q000097:blood / Q000494:pharmacology
D016023:Integrins; D009842:Oligopeptides; D019030:Receptors, Vitronectin; D019096:Vitronectin; C064660:integrin alphaVbeta5; C047981:arginyl-glycyl-aspartic acid; D014568:Urokinase-Type Plasminogen Activator