Regulation of urokinase receptor proteolytic function by the tetraspanin CD82.
The high affinity interaction between the urokinase-type plasminogen activator (uPA) and its glycolipid-anchored cellular receptor (uPAR) promotes plasminogen activation and the efficient generation of pericellular proteolytic activity. We demonstrate here that expression of the tetraspanin CD82/KAI1 (a tumor metastasis suppressor) leads to a profound effect on uPAR function. Pericellular plasminogen activation was reduced by approximately 50-fold in the presence of CD82, although levels of components of the plasminogen activation system were unchanged. uPAR was present on the cell surface and molecularly intact, but radioligand binding analysis with uPA and anti-uPAR antibodies revealed that it was in a previously undetected cryptic form unable to bind uPA. This was not due to direct interactions between uPAR and CD82, as they neither co-localized on the cell surface nor could be co-immunoprecipitated. However, expression of CD82 led to a redistribution of uPAR to focal adhesions, where it was shown by double immunofluorescence labeling to co-localize with the integrin alpha(5)beta(1), which was also redistributed in the presence of CD82. Co-immunoprecipitation experiments showed that, in the presence of CD82, uPAR preferentially formed stable associations with alpha(5)beta(1), but not with a variety of other integrins, including alpha(3)beta(1). These data suggest that CD82 inhibits the proteolytic function of uPAR indirectly, directing uPAR and alpha(5)beta(1) to focal adhesions and promoting their association with a resultant loss of uPA binding. This represents a novel mechanism whereby tetraspanins, integrins, and uPAR, systems involved in cell adhesion and migration, cooperate to regulate pericellular proteolytic activity and may suggest a mechanism for the tumor-suppressive effects of CD82/KAI1.
Bass
Rosemary
R
School of Biological Science, University of East Anglia, Norwich NR4 7TJ, United Kingdom.
Werner
Finn
F
Odintsova
Elena
E
Sugiura
Tsuyoshi
T
Berditchevski
Fedor
F
Ellis
Vincent
V
D015703:Antigens, CD / Q000096:biosynthesis / Q000737:chemistry* / Q000502:physiology*; D019921:Biotinylation; D002448:Cell Adhesion; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D002465:Cell Movement; D003432:Cross-Linking Reagents / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D022001:Focal Adhesions / Q000378:metabolism; D005732:Gangliosides / Q000494:pharmacology; D006801:Humans; D007150:Immunohistochemistry; D047468:Immunoprecipitation; D039201:Integrin alpha3beta1 / Q000378:metabolism; D039081:Integrin alpha5beta1 / Q000378:metabolism; D016023:Integrins / Q000378:metabolism; D051861:Kangai-1 Protein; D042361:Mammary Glands, Human / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000096:biosynthesis / Q000737:chemistry* / Q000502:physiology*; D008856:Microscopy, Fluorescence; D010958:Plasminogen / Q000737:chemistry; D010960:Plasminogen Activators / Q000737:chemistry; D011485:Protein Binding; D011518:Proto-Oncogene Proteins / Q000096:biosynthesis / Q000737:chemistry* / Q000502:physiology*; D011956:Receptors, Cell Surface / Q000737:chemistry / Q000378:metabolism / Q000502:physiology*; D055293:Receptors, Urokinase Plasminogen Activator; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors
D015703:Antigens, CD; C497688:CD82 protein, human; D003432:Cross-Linking Reagents; D005732:Gangliosides; D039201:Integrin alpha3beta1; D039081:Integrin alpha5beta1; D016023:Integrins; D051861:Kangai-1 Protein; D008562:Membrane Glycoproteins; C524898:PLAUR protein, human; D011518:Proto-Oncogene Proteins; D011956:Receptors, Cell Surface; D055293:Receptors, Urokinase Plasminogen Activator; D010958:Plasminogen; D010960:Plasminogen Activators