Urokinase receptors promote beta1 integrin function through interactions with integrin alpha3beta1.
The urokinase receptor (uPAR) is linked to cellular migration through its capacity to promote pericellular proteolysis, regulate integrin function, and mediate cell signaling in response to urokinase (uPA) binding. The mechanisms for these activities remain incompletely defined, although uPAR was recently identified as a cis-acting ligand for the beta2 integrin CD11b/CD18 (Mac-1). Here we show that a major beta1 integrin partner for uPAR/uPA signaling is alpha3. In uPAR-transfected 293 cells uPAR complexed (>90%) with alpha3beta1 and antibodies to alpha3 blocked uPAR-dependent vitronectin (Vn) adhesion. Soluble uPAR bound to recombinant alpha3beta1 in a uPA-dependent manner (K(d) < 20 nM) and binding was blocked by a 17-mer alpha3beta1 integrin peptide (alpha325) homologous to the CD11b uPAR-binding site. uPAR colocalized with alpha3beta1 in MDA-MB-231 cells and uPA (1 nM) enhanced spreading and focal adhesion kinase phosphorylation on fibronectin (Fn) or collagen type I (Col) in a pertussis toxin- and alpha325-sensitive manner. A critical role of alpha3beta1 in uPA signaling was verified by studies of epithelial cells from alpha3-deficient mice. Thus, uPAR preferentially complexes with alpha3beta1, promoting direct (Vn) and indirect (Fn, Col) pathways of cell adhesion, the latter a heterotrimeric G protein-dependent mechanism of signaling between alpha3beta1 and other beta1 integrins.
Wei
Y
Y
Respiratory Division, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115, USA.
Eble
J A
JA
Wang
Z
Z
Kreidberg
J A
JA
Chapman
H A
HA
D001665:Binding Sites / Q000502:physiology; D018821:CD18 Antigens; D002460:Cell Line; D002465:Cell Movement / Q000187:drug effects / Q000502:physiology*; D003094:Collagen / Q000378:metabolism; D005353:Fibronectins / Q000378:metabolism; D020962:Heterotrimeric GTP-Binding Proteins / Q000737:chemistry; D006801:Humans; D039201:Integrin alpha3beta1; D019012:Integrin beta1 / Q000378:metabolism*; D016023:Integrins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D016177:Macrophage-1 Antigen / Q000235:genetics / Q000378:metabolism; D011233:Precipitin Tests; D011956:Receptors, Cell Surface / Q000378:metabolism*; D055293:Receptors, Urokinase Plasminogen Activator; D011994:Recombinant Proteins / Q000378:metabolism; D017385:Sequence Homology; D015398:Signal Transduction; D014407:Tumor Cells, Cultured; D014568:Urokinase-Type Plasminogen Activator / Q000378:metabolism* / Q000494:pharmacology; D019061:src-Family Kinases / Q000737:chemistry
D018821:CD18 Antigens; D005353:Fibronectins; D039201:Integrin alpha3beta1; D019012:Integrin beta1; D016023:Integrins; D016177:Macrophage-1 Antigen; C524898:PLAUR protein, human; C524899:Plaur protein, mouse; D011956:Receptors, Cell Surface; D055293:Receptors, Urokinase Plasminogen Activator; D011994:Recombinant Proteins; D003094:Collagen; D019061:src-Family Kinases; D014568:Urokinase-Type Plasminogen Activator; D020962:Heterotrimeric GTP-Binding Proteins