Skip to main content
PMC home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1996 Sep 2;134(6):1563–1571. doi: 10.1083/jcb.134.6.1563

Is plasminogen activator inhibitor-1 the molecular switch that governs urokinase receptor-mediated cell adhesion and release?

PMCID: PMC2120994  PMID: 8830783

Abstract

Induction of the urokinase type plasminogen activator receptor (uPAR) promotes cell adhesion through its interaction with vitronectin (VN) in the extracellular matrix, and facilitates cell migration and invasion by localizing uPA to the cell surface. We provide evidence that this balance between cell adhesion and cell detachment is governed by PA inhibitor-1 (PAI-1). First, we demonstrate that uPAR and PAI-1 bind to the same site in VN (i.e., the amino-terminal somatomedin B domain; SMB), and that PAI-1 competes with uPAR for binding to SMB. Domain swapping and mutagenesis studies indicate that the uPAR-binding sequence is located within the central region of the SMB domain, a region previously shown to contain the PAI-1-binding motif. Second, we show that PAI-1 dissociates bound VN from uPAR and detaches U937 cells from their VN substratum. This PAI-1 mediated release of cells from VN appears to occur independently of its ability to function as a protease inhibitor, and may help to explain why high PAI-1 levels indicate a poor prognosis for many cancers. Finally, we show that uPA can rapidly reverse this effect of PAI-1. Taken together, these results suggest a dynamic regulatory role for PAI-1 and uPA in uPAR-mediated cell adhesion and release.

Full Text

The Full Text of this article is available as a PDF (1.5 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Andreasen P. A., Nielsen L. S., Kristensen P., Grøndahl-Hansen J., Skriver L., Danø K. Plasminogen activator inhibitor from human fibrosarcoma cells binds urokinase-type plasminogen activator, but not its proenzyme. J Biol Chem. 1986 Jun 15;261(17):7644–7651. [PubMed] [Google Scholar]
  2. Behrendt N., Rønne E., Danø K. The structure and function of the urokinase receptor, a membrane protein governing plasminogen activation on the cell surface. Biol Chem Hoppe Seyler. 1995 May;376(5):269–279. [PubMed] [Google Scholar]
  3. Bernfield M., Hinkes M. T., Gallo R. L. Developmental expression of the syndecans: possible function and regulation. Dev Suppl. 1993:205–212. [PubMed] [Google Scholar]
  4. Boukerche H., Benchaibi M., Berthier-Vergnes O., Lizard G., Bailly M., Bailly M., McGregor J. L. Two human melanoma cell-line variants with enhanced in vivo tumor growth and metastatic capacity do not express the beta 3 integrin subunit. Eur J Biochem. 1994 Mar 1;220(2):485–491. doi: 10.1111/j.1432-1033.1994.tb18647.x. [DOI] [PubMed] [Google Scholar]
  5. Chen W. T. Membrane proteases: roles in tissue remodeling and tumour invasion. Curr Opin Cell Biol. 1992 Oct;4(5):802–809. doi: 10.1016/0955-0674(92)90103-j. [DOI] [PubMed] [Google Scholar]
  6. Ciambrone G. J., McKeown-Longo P. J. Vitronectin regulates the synthesis and localization of urokinase-type plasminogen activator in HT-1080 cells. J Biol Chem. 1992 Jul 5;267(19):13617–13622. [PubMed] [Google Scholar]
  7. Cohen R. L., Xi X. P., Crowley C. W., Lucas B. K., Levinson A. D., Shuman M. A. Effects of urokinase receptor occupancy on plasmin generation and proteolysis of basement membrane by human tumor cells. Blood. 1991 Jul 15;78(2):479–487. [PubMed] [Google Scholar]
  8. Cubellis M. V., Andreasen P., Ragno P., Mayer M., Danø K., Blasi F. Accessibility of receptor-bound urokinase to type-1 plasminogen activator inhibitor. Proc Natl Acad Sci U S A. 1989 Jul;86(13):4828–4832. doi: 10.1073/pnas.86.13.4828. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Cubellis M. V., Wun T. C., Blasi F. Receptor-mediated internalization and degradation of urokinase is caused by its specific inhibitor PAI-1. EMBO J. 1990 Apr;9(4):1079–1085. doi: 10.1002/j.1460-2075.1990.tb08213.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Danø K., Andreasen P. A., Grøndahl-Hansen J., Kristensen P., Nielsen L. S., Skriver L. Plasminogen activators, tissue degradation, and cancer. Adv Cancer Res. 1985;44:139–266. doi: 10.1016/s0065-230x(08)60028-7. [DOI] [PubMed] [Google Scholar]
  11. Declerck P. J., De Mol M., Alessi M. C., Baudner S., Pâques E. P., Preissner K. T., Müller-Berghaus G., Collen D. Purification and characterization of a plasminogen activator inhibitor 1 binding protein from human plasma. Identification as a multimeric form of S protein (vitronectin). J Biol Chem. 1988 Oct 25;263(30):15454–15461. [PubMed] [Google Scholar]
  12. Del Vecchio S., Stoppelli M. P., Carriero M. V., Fonti R., Massa O., Li P. Y., Botti G., Cerra M., D'Aiuto G., Esposito G. Human urokinase receptor concentration in malignant and benign breast tumors by in vitro quantitative autoradiography: comparison with urokinase levels. Cancer Res. 1993 Jul 1;53(13):3198–3206. [PubMed] [Google Scholar]
  13. Delbaldo C., Cunningham M., Vassalli J. D., Sappino A. P. Plasmin-catalyzed proteolysis in colorectal neoplasia. Cancer Res. 1995 Oct 15;55(20):4688–4695. [PubMed] [Google Scholar]
  14. Deng G., Royle G., Seiffert D., Loskutoff D. J. The PAI-1/vitronectin interaction: two cats in a bag? Thromb Haemost. 1995 Jul;74(1):66–70. [PubMed] [Google Scholar]
  15. Deng G., Royle G., Wang S., Crain K., Loskutoff D. J. Structural and functional analysis of the plasminogen activator inhibitor-1 binding motif in the somatomedin B domain of vitronectin. J Biol Chem. 1996 May 31;271(22):12716–12723. doi: 10.1074/jbc.271.22.12716. [DOI] [PubMed] [Google Scholar]
  16. Estreicher A., Mühlhauser J., Carpentier J. L., Orci L., Vassalli J. D. The receptor for urokinase type plasminogen activator polarizes expression of the protease to the leading edge of migrating monocytes and promotes degradation of enzyme inhibitor complexes. J Cell Biol. 1990 Aug;111(2):783–792. doi: 10.1083/jcb.111.2.783. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Gladson C. L., Pijuan-Thompson V., Olman M. A., Gillespie G. Y., Yacoub I. Z. Up-regulation of urokinase and urokinase receptor genes in malignant astrocytoma. Am J Pathol. 1995 May;146(5):1150–1160. [PMC free article] [PubMed] [Google Scholar]
  18. Goodson R. J., Doyle M. V., Kaufman S. E., Rosenberg S. High-affinity urokinase receptor antagonists identified with bacteriophage peptide display. Proc Natl Acad Sci U S A. 1994 Jul 19;91(15):7129–7133. doi: 10.1073/pnas.91.15.7129. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Grøndahl-Hansen J., Christensen I. J., Rosenquist C., Brünner N., Mouridsen H. T., Danø K., Blichert-Toft M. High levels of urokinase-type plasminogen activator and its inhibitor PAI-1 in cytosolic extracts of breast carcinomas are associated with poor prognosis. Cancer Res. 1993 Jun 1;53(11):2513–2521. [PubMed] [Google Scholar]
  20. Gumbiner B. M. Cell adhesion: the molecular basis of tissue architecture and morphogenesis. Cell. 1996 Feb 9;84(3):345–357. doi: 10.1016/s0092-8674(00)81279-9. [DOI] [PubMed] [Google Scholar]
  21. HE C. S., Wilhelm S. M., Pentland A. P., Marmer B. L., Grant G. A., Eisen A. Z., Goldberg G. I. Tissue cooperation in a proteolytic cascade activating human interstitial collagenase. Proc Natl Acad Sci U S A. 1989 Apr;86(8):2632–2636. doi: 10.1073/pnas.86.8.2632. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Hauptmann S., Denkert C., Löhrke H., Tietze L., Ott S., Klosterhalfen B., Mittermayer C. Integrin expression on colorectal tumor cells growing as monolayers, as multicellular tumor spheroids, or in nude mice. Int J Cancer. 1995 Jun 9;61(6):819–825. doi: 10.1002/ijc.2910610613. [DOI] [PubMed] [Google Scholar]
  23. Heiss M. M., Babic R., Allgayer H., Gruetzner K. U., Jauch K. W., Loehrs U., Schildberg F. W. Tumor-associated proteolysis and prognosis: new functional risk factors in gastric cancer defined by the urokinase-type plasminogen activator system. J Clin Oncol. 1995 Aug;13(8):2084–2093. doi: 10.1200/JCO.1995.13.8.2084. [DOI] [PubMed] [Google Scholar]
  24. Howlett A. R., Bailey N., Damsky C., Petersen O. W., Bissell M. J. Cellular growth and survival are mediated by beta 1 integrins in normal human breast epithelium but not in breast carcinoma. J Cell Sci. 1995 May;108(Pt 5):1945–1957. doi: 10.1242/jcs.108.5.1945. [DOI] [PubMed] [Google Scholar]
  25. Huarte J., Belin D., Vassalli J. D. Plasminogen activator in mouse and rat oocytes: induction during meiotic maturation. Cell. 1985 Dec;43(2 Pt 1):551–558. doi: 10.1016/0092-8674(85)90184-9. [DOI] [PubMed] [Google Scholar]
  26. Hynes R. O., Lander A. D. Contact and adhesive specificities in the associations, migrations, and targeting of cells and axons. Cell. 1992 Jan 24;68(2):303–322. doi: 10.1016/0092-8674(92)90472-o. [DOI] [PubMed] [Google Scholar]
  27. Jenne D., Stanley K. K. Molecular cloning of S-protein, a link between complement, coagulation and cell-substrate adhesion. EMBO J. 1985 Dec 1;4(12):3153–3157. doi: 10.1002/j.1460-2075.1985.tb04058.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Kaufman S. E., Brown S., Stauber G. B. Characterization of ligand binding to immobilized biotinylated extracellular domains of three growth factor receptors. Anal Biochem. 1993 Jun;211(2):261–266. doi: 10.1006/abio.1993.1267. [DOI] [PubMed] [Google Scholar]
  29. Knudsen B. S., Harpel P. C., Nachman R. L. Plasminogen activator inhibitor is associated with the extracellular matrix of cultured bovine smooth muscle cells. J Clin Invest. 1987 Oct;80(4):1082–1089. doi: 10.1172/JCI113164. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Kobayashi H., Fujishiro S., Terao T. Impact of urokinase-type plasminogen activator and its inhibitor type 1 on prognosis in cervical cancer of the uterus. Cancer Res. 1994 Dec 15;54(24):6539–6548. [PubMed] [Google Scholar]
  31. Kruithof E. K. Plasminogen activator inhibitors--a review. Enzyme. 1988;40(2-3):113–121. doi: 10.1159/000469153. [DOI] [PubMed] [Google Scholar]
  32. Kuhn W., Pache L., Schmalfeldt B., Dettmar P., Schmitt M., Jänicke F., Graeff H. Urokinase (uPA) and PAI-1 predict survival in advanced ovarian cancer patients (FIGO III) after radical surgery and platinum-based chemotherapy. Gynecol Oncol. 1994 Dec;55(3 Pt 1):401–409. doi: 10.1006/gyno.1994.1313. [DOI] [PubMed] [Google Scholar]
  33. Liu G., Shuman M. A., Cohen R. L. Co-expression of urokinase, urokinase receptor and PAI-1 is necessary for optimum invasiveness of cultured lung cancer cells. Int J Cancer. 1995 Feb 8;60(4):501–506. doi: 10.1002/ijc.2910600413. [DOI] [PubMed] [Google Scholar]
  34. Mignatti P., Rifkin D. B. Biology and biochemistry of proteinases in tumor invasion. Physiol Rev. 1993 Jan;73(1):161–195. doi: 10.1152/physrev.1993.73.1.161. [DOI] [PubMed] [Google Scholar]
  35. Mimuro J., Loskutoff D. J. Binding of type 1 plasminogen activator inhibitor to the extracellular matrix of cultured bovine endothelial cells. J Biol Chem. 1989 Mar 25;264(9):5058–5063. [PubMed] [Google Scholar]
  36. Nekarda H., Schmitt M., Ulm K., Wenninger A., Vogelsang H., Becker K., Roder J. D., Fink U., Siewert J. R. Prognostic impact of urokinase-type plasminogen activator and its inhibitor PAI-1 in completely resected gastric cancer. Cancer Res. 1994 Jun 1;54(11):2900–2907. [PubMed] [Google Scholar]
  37. Nesbit M., Herlyn M. Adhesion receptors in human melanoma progression. Invasion Metastasis. 1994;14(1-6):131–146. [PubMed] [Google Scholar]
  38. Pappot H., Gårdsvoll H., Rømer J., Pedersen A. N., Grøndahl-Hansen J., Pyke C., Brünner N. Plasminogen activator inhibitor type 1 in cancer: therapeutic and prognostic implications. Biol Chem Hoppe Seyler. 1995 May;376(5):259–267. doi: 10.1515/bchm3.1995.376.5.259. [DOI] [PubMed] [Google Scholar]
  39. Pedersen H., Brünner N., Francis D., Osterlind K., Rønne E., Hansen H. H., Danø K., Grøndahl-Hansen J. Prognostic impact of urokinase, urokinase receptor, and type 1 plasminogen activator inhibitor in squamous and large cell lung cancer tissue. Cancer Res. 1994 Sep 1;54(17):4671–4675. [PubMed] [Google Scholar]
  40. Ploug M., Ellis V., Danø K. Ligand interaction between urokinase-type plasminogen activator and its receptor probed with 8-anilino-1-naphthalenesulfonate. Evidence for a hydrophobic binding site exposed only on the intact receptor. Biochemistry. 1994 Aug 2;33(30):8991–8997. doi: 10.1021/bi00196a017. [DOI] [PubMed] [Google Scholar]
  41. Ploug M., Rønne E., Behrendt N., Jensen A. L., Blasi F., Danø K. Cellular receptor for urokinase plasminogen activator. Carboxyl-terminal processing and membrane anchoring by glycosyl-phosphatidylinositol. J Biol Chem. 1991 Jan 25;266(3):1926–1933. [PubMed] [Google Scholar]
  42. Plow E. F., Herren T., Redlitz A., Miles L. A., Hoover-Plow J. L. The cell biology of the plasminogen system. FASEB J. 1995 Jul;9(10):939–945. doi: 10.1096/fasebj.9.10.7615163. [DOI] [PubMed] [Google Scholar]
  43. Prendergast G. C., Cole M. D. Posttranscriptional regulation of cellular gene expression by the c-myc oncogene. Mol Cell Biol. 1989 Jan;9(1):124–134. doi: 10.1128/mcb.9.1.124. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Quax P. H., van Muijen G. N., Weening-Verhoeff E. J., Lund L. R., Danø K., Ruiter D. J., Verheijen J. H. Metastatic behavior of human melanoma cell lines in nude mice correlates with urokinase-type plasminogen activator, its type-1 inhibitor, and urokinase-mediated matrix degradation. J Cell Biol. 1991 Oct;115(1):191–199. doi: 10.1083/jcb.115.1.191. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Roldan A. L., Cubellis M. V., Masucci M. T., Behrendt N., Lund L. R., Danø K., Appella E., Blasi F. Cloning and expression of the receptor for human urokinase plasminogen activator, a central molecule in cell surface, plasmin dependent proteolysis. EMBO J. 1990 Feb;9(2):467–474. doi: 10.1002/j.1460-2075.1990.tb08132.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Roussel E., Gingras M. C., Ro J. Y., Branch C., Roth J. A. Loss of alpha 1 beta 1 and reduced expression of other beta 1 integrins and CAM in lung adenocarcinoma compared with pneumocytes. J Surg Oncol. 1994 Jul;56(3):198–208. doi: 10.1002/jso.2930560315. [DOI] [PubMed] [Google Scholar]
  47. Ruoslahti E. Fibronectin and its alpha 5 beta 1 integrin receptor in malignancy. Invasion Metastasis. 1994;14(1-6):87–97. [PubMed] [Google Scholar]
  48. Salonen E. M., Vaheri A., Pöllänen J., Stephens R., Andreasen P., Mayer M., Danø K., Gailit J., Ruoslahti E. Interaction of plasminogen activator inhibitor (PAI-1) with vitronectin. J Biol Chem. 1989 Apr 15;264(11):6339–6343. [PubMed] [Google Scholar]
  49. Seiffert D., Ciambrone G., Wagner N. V., Binder B. R., Loskutoff D. J. The somatomedin B domain of vitronectin. Structural requirements for the binding and stabilization of active type 1 plasminogen activator inhibitor. J Biol Chem. 1994 Jan 28;269(4):2659–2666. [PubMed] [Google Scholar]
  50. Seiffert D., Loskutoff D. J. Evidence that type 1 plasminogen activator inhibitor binds to the somatomedin B domain of vitronectin. J Biol Chem. 1991 Feb 15;266(5):2824–2830. [PubMed] [Google Scholar]
  51. Seiffert D., Loskutoff D. J. Kinetic analysis of the interaction between type 1 plasminogen activator inhibitor and vitronectin and evidence that the bovine inhibitor binds to a thrombin-derived amino-terminal fragment of bovine vitronectin. Biochim Biophys Acta. 1991 May 30;1078(1):23–30. doi: 10.1016/0167-4838(91)90087-g. [DOI] [PubMed] [Google Scholar]
  52. Sier C. F., Vloedgraven H. J., Ganesh S., Griffioen G., Quax P. H., Verheijen J. H., Dooijewaard G., Welvaart K., van de Velde C. J., Lamers C. B. Inactive urokinase and increased levels of its inhibitor type 1 in colorectal cancer liver metastasis. Gastroenterology. 1994 Nov;107(5):1449–1456. doi: 10.1016/0016-5085(94)90549-5. [DOI] [PubMed] [Google Scholar]
  53. Stratton-Thomas J. R., Min H. Y., Kaufman S. E., Chiu C. Y., Mullenbach G. T., Rosenberg S. Yeast expression and phagemid display of the human urokinase plasminogen activator epidermal growth factor-like domain. Protein Eng. 1995 May;8(5):463–470. doi: 10.1093/protein/8.5.463. [DOI] [PubMed] [Google Scholar]
  54. Vassalli J. D., Sappino A. P., Belin D. The plasminogen activator/plasmin system. J Clin Invest. 1991 Oct;88(4):1067–1072. doi: 10.1172/JCI115405. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Waltz D. A., Chapman H. A. Reversible cellular adhesion to vitronectin linked to urokinase receptor occupancy. J Biol Chem. 1994 May 20;269(20):14746–14750. [PubMed] [Google Scholar]
  56. Waltz D. A., Sailor L. Z., Chapman H. A. Cytokines induce urokinase-dependent adhesion of human myeloid cells. A regulatory role for plasminogen activator inhibitors. J Clin Invest. 1993 Apr;91(4):1541–1552. doi: 10.1172/JCI116360. [DOI] [PMC free article] [PubMed] [Google Scholar]
  57. Wei Y., Waltz D. A., Rao N., Drummond R. J., Rosenberg S., Chapman H. A. Identification of the urokinase receptor as an adhesion receptor for vitronectin. J Biol Chem. 1994 Dec 23;269(51):32380–32388. [PubMed] [Google Scholar]
  58. Wiman B., Almquist A., Sigurdardottir O., Lindahl T. Plasminogen activator inhibitor 1 (PAI) is bound to vitronectin in plasma. FEBS Lett. 1988 Dec 19;242(1):125–128. doi: 10.1016/0014-5793(88)80999-2. [DOI] [PubMed] [Google Scholar]
  59. Yatohgo T., Izumi M., Kashiwagi H., Hayashi M. Novel purification of vitronectin from human plasma by heparin affinity chromatography. Cell Struct Funct. 1988 Aug;13(4):281–292. doi: 10.1247/csf.13.281. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES