Thromb Haemost 2013; 109(02): 309-318
DOI: 10.1160/TH12-08-0546
Cellular Proteolysis and Oncology
Schattauer GmbH

uPAR regulates pericellular proteolysis through a mechanism involving integrins and fMLF-receptors

Nunzia Montuori*
1   Department of Cellular and Molecular Biology and Pathology, “Federico II” University, Naples, Italy
,
Vincenzo Cosimato*
2   Department of Chemistry and Biology, University of Salerno, Fisciano (Salerno), Italy
,
Loredana Rinaldi
2   Department of Chemistry and Biology, University of Salerno, Fisciano (Salerno), Italy
,
Vincenza Elena Anna Rea
1   Department of Cellular and Molecular Biology and Pathology, “Federico II” University, Naples, Italy
,
Daniela Alfano§
2   Department of Chemistry and Biology, University of Salerno, Fisciano (Salerno), Italy
,
Pia Ragno
2   Department of Chemistry and Biology, University of Salerno, Fisciano (Salerno), Italy
› Author Affiliations
Further Information

Publication History

Received: 07 August 2012

Accepted after major revision: 09 November 2012

Publication Date:
29 November 2017 (online)

Summary

The expression of the urokinase-type plasminogen activator (uPA) and its receptor (uPAR) can be regulated by several hormones, cytokines, and tumour promoters. uPAR is a glycosyl-phosphatidyl inositol (GPI)- linked cell-surface protein; however, it is capable to transduce signals inside the cell by interacting with other cell-surface proteins, such as integrins and G-protein coupled (GPC) receptors. We previously reported that uPAR cell-surface expression can be positively regulated by its ligand, uPA, independently of its proteolytic activity. We now demonstrate that uPAR overexpression induces or increases uPA secretion both in uPAR-negative and in uPAR-expressing cells. Accordingly, uPAR depletion impairs uPA expression in cells which constitutively express both uPA and its receptor. uPAR exerts its regulatory effect through the activation of the ERK mitogen-activated protein kinases (MAPKs), whereas the p-38 MAPK is not involved. Overexpression of truncated forms of uPAR, lacking the N-terminal domain (DI) and not able to interact with membrane co-receptors, failed to increase uPA expression. Inhibition of uPAR-integrin interaction by the specific P-25 peptide, as well as Gi-protein inhibition by cholera pertussin toxin or depletion of the GPC receptors for fMLF (fMLF-Rs) also impaired uPAR capability to regulate uPA expression. These findings demonstrate that uPAR, whose expression is regulated by uPA, can, in turn, regulate uPA expression through a mechanism involving its functional interaction with integrins and fMLF-Rs.

* These authors contributed equally to this work.


§ Current address: Institute of Genetics and Biophysics Adriano Buzzati- Traverso, Consiglio Nazionale delle Ricerche (CNR), Naples, Italy.


 
  • References

  • 1 Ragno P. The urokinase receptor: a ligand or a receptor? Story of a sociable molecule. Cell Mol Life Sci 2006; 63: 1028-1037.
  • 2 Ellis V, Scully MF, Kakkar VV. Plasminogen activation initiated by single-chain urokinase-type plasminogen activator. Potentiation by U937 monocytes. J Biol Chem 1989; 264: 2185-2188.
  • 3 Montuori N, Ragno P. Multiple activities of a multifaceted receptor: roles of cleaved and soluble uPAR. Front Biosci 2009; 14: 2494-2503.
  • 4 Fazioli F, Resnati M, Sidenius N. et al. A urokinase-sensitive region of the human urokinase receptor is responsible for its chemotactic activity. EMBO J 1997; 16: 7279-7286.
  • 5 Resnati M, Pallavicini I, Wang JM. et al. The fibrinolytic receptor for urokinase activates the G protein-coupled chemotactic receptor FPRL1/LXA4R. Proc Natl Acad Sci USA 2002; 99: 1359-1364.
  • 6 Huang J, Chen K, Gong W. et al. G-protein coupled chemoattractant receptors and cancer. Front Biosci 2008; 13: 3352-3363.
  • 7 Montuori N, Carriero MV, Salzano S. et al. The cleavage of the urokinase receptor regulates its multiple functions. J Biol Chem 2002; 277: 46932-46939.
  • 8 de Paulis A, Montuori N, Prevete N. et al. Urokinase induces basophil chemotaxis through a urokinase receptor epitope that is an endogenous ligand for formyl peptide receptor-like 1 and -like 2. J Immunol 2004; 173: 5739-5748.
  • 9 D’Alessio S, Blasi F. The urokinase receptor as an entertainer of signal transduction. Front Biosci 2009; 14: 4575-4587.
  • 10 Smith HW, Marshall CJ. Regulation of cell signalling by uPAR. Nat Rev Mol Cell Biol 2010; 11: 23-36.
  • 11 Blasi F, Carmeliet P. uPAR: a versatile signalling orchestrator. Nat Rev Mol Cell Biol 2002; 03: 932-943.
  • 12 Montuori N, Visconte V, Rossi G. et al. Soluble and cleaved forms of the urokinase-receptor: degradation products or active molecules?. Thromb Haemost 2005; 93: 192-198.
  • 13 Montuori N, Rossi G, Ragno P. Post-transcriptional regulation of gene expression in the plasminogen activation system. Biol Chem 2002; 383: 47-53.
  • 14 Montuori N, Salzano S, Rossi G. et al. Urokinase-type plasminogen activator up-regulates the expression of its cellular receptor. FEBS Lett 2000; 476: 166-170.
  • 15 Montuori N, Mattiello A, Mancini A. et al. Urokinase-type plasminogen activator up-regulates the expression of its cellular receptor through a post-transcriptional mechanism. FEBS Lett 2001; 508: 379-384.
  • 16 Montuori N, Mattiello A, Mancini A. et al. Urokinase-mediated posttranscriptional regulation of urokinase-receptor expression in non small cell lung carcinoma. Int J Cancer 2003; 105: 353-360.
  • 17 R∅nne E, Behrendt N, Ellis V. et al. Cell-induced potentiation of the plasminogen activation system is abolished by a monoclonal antibody that recognizes the NH2-terminal domain of the urokinase receptor. FEBS Lett 1991; 288: 233-236.
  • 18 Roldan AL, Cubellis MV, Masucci MT. et al. Cloning and expression of the receptor for human urokinase plasminogen activator, a central molecule in cell surface, plasmin dependent proteolysis. EMBO J 1990; 09: 467-474.
  • 19 Vassalli JD, Dayer JM, Wohlwend A. et al. Concomitant secretion of prourokinase and of a plasminogen activator-specific inhibitor by cultured human monocytes-macrophages. J Exp Med 1984; 159: 1653-1668.
  • 20 Ragno P, Estreicher A, Gos A. et al. Polarized secretion of urokinase-type plasminogen activator by epithelial cells. Exp Cell Res 1992; 203: 236-243.
  • 21 Wei Y, Lukashev M, Simon DI. et al. Regulation of integrin function by the urokinase receptor. Science 1996; 273: 1551-1555.
  • 22 Wei Y, Waltz DA, Rao N. et al. Identification of the urokinase receptor as an adhesion receptor for vitronectin. J Biol Chem 1994; 269: 32380-32388.
  • 23 Madsen CD, Ferraris GM, Andolfo A. et al. uPAR-induced cell adhesion and migration: vitronectin provides the key. J Cell Biol 2007; 177: 927-939.
  • 24 Besser D, Presta M, Nagamine Y. Elucidation of a signalling pathway induced by FGF-2 leading to uPA gene expression in NIH 3T3 fibroblasts. Cell Growth Differ 1995; 06: 1009-1017.
  • 25 Silberman S, Janulis M, Schultz RM. Characterization of downstream Ras signals that induce alternative protease-dependent invasive phenotypes. J Biol Chem 1997; 272: 5927-5935.
  • 26 Huang S, New L, Pan Z. et al. Urokinase plasminogen activator/urokinase-specific surface receptor expression and matrix invasion by breast cancer cells requires constitutive p38alpha mitogen-activated protein kinase activity. J Biol Chem 2000; 275: 12266-12272.
  • 27 Montero L, Nagamine Y. Regulation by p38 mitogen-activated protein kinase of adenylate- and uridylate-rich element-mediated urokinase-type plasminogen activator (uPA) messenger RNA stability and uPA-dependent in vitro cell invasion. Cancer Res 1999; 59: 5286-5293.
  • 28 Gyetko MR, Todd 3rd RF, Wilkinson CC. et al. The urokinase receptor is required for human monocyte chemotaxis in vitro. J Clin Invest 1994; 93: 1380-1387.
  • 29 Schreiber RE, Prossnitz ER, Ye RD. et al. Reconstitution of recombinant N-formyl chemotactic peptide receptor with G protein. J Leukoc Biol 1993; 53: 470-474.
  • 30 Wei Y, Eble JA, Wang Z. et al. Urokinase receptors promote beta1 integrin function through interactions with integrin alpha3beta1. Mol Biol Cell 2001; 12: 2975-2986.
  • 31 Zannetti A, Del Vecchio S, Carriero MV. et al. Coordinate up-regulation of Sp1 DNA-binding activity and urokinase receptor expression in breast carcinoma. Cancer Res 2000; 60: 1546-1551.
  • 32 Gargiulo L, Longanesi-Cattani I, Bifulco K. et al. Cross-talk between fMLP and vitronectin receptors triggered by urokinase receptor-derived SRSRY peptide. J Biol Chem 2005; 280: 25225-25232.
  • 33 Chigaev A, Waller A, Amit O. et al. Real-time analysis of conformation-sensitive antibody binding provides new insights into integrin conformational regulation. J Biol Chem 2009; 284: 14337-14346.
  • 34 Ghosh S, Johnson JJ, Sen R. et al. Functional relevance of urinary-type plasminogen activator receptor-alpha3beta1 integrin association in proteinase regulatory pathways. J Biol Chem 2006; 281: 13021-13029.
  • 35 Hildenbrand R, Schaaf A, Dorn-Beineke A. et al. Tumor stroma is the predominant uPA-, uPAR-, PAI-1-expressing tissue in human breast cancer: prognostic impact. Histol Histopathol 2009; 24: 869-877.
  • 36 GrØndahl-Hansen J, Ralfkiaer E, Kirkeby LT. et al. Localization of urokinase-type plasminogen activator in stromal cells in adenocarcinomas of the colon in humans. Am J Pathol 1991; 138: 111-117.