Transcriptional Regulation of the Murine Urokinase-type Plasminogen Activator Gene in Skeletal Myoblasts

Francesc Miralles; Inés Ibáñez-Tallon; Maribel Parra; Massimo Crippa; Francesco Blasi; Daniel Besser; Yoshikuni Nagamine; Pura Muñoz-Cánoves

doi:10.1055/s-0037-1614569

Thrombosis and Haemostasis, Inhaltsverzeichnis

Thromb Haemost 1999; 81(05): 767-774
DOI: 10.1055/s-0037-1614569

Rapid Communication

Schattauer GmbH

Transcriptional Regulation of the Murine Urokinase-type Plasminogen Activator Gene in Skeletal Myoblasts

Francesc Miralles

¹From the Institut de Recerca Oncològica (IRO), Barcelona, Spain

,

Inés Ibáñez-Tallon

²Laboratory of Molecular Genetics, DIBIT-H. S. Raffaele, Milan, Italy

,

Maribel Parra

¹From the Institut de Recerca Oncològica (IRO), Barcelona, Spain

,

Massimo Crippa

²Laboratory of Molecular Genetics, DIBIT-H. S. Raffaele, Milan, Italy

,

Francesco Blasi

²Laboratory of Molecular Genetics, DIBIT-H. S. Raffaele, Milan, Italy

,

Daniel Besser

³Friedrich Miescher Institut, Basel, Switzerland

,

Yoshikuni Nagamine

³Friedrich Miescher Institut, Basel, Switzerland

,

Pura Muñoz-Cánoves

¹From the Institut de Recerca Oncològica (IRO), Barcelona, Spain

› Institutsangaben

Abstract

Summary

We have previously shown that urokinase-type plasminogen activator (uPA) is highly expressed in murine C2C12 myoblasts and that antibodies against uPA are able to block both myoblast fusion and differentiation. Here we show the characterization of cis-acting elements in the mouse uPA promoter in vitro which are involved in uPA gene expression in C2C12 myoblast cells. DNase I hypersensitive (HS) site analysis revealed the presence of three HS sites in myoblasts. Deletion analysis of stably transfected uPA-promoter constructs revealed that at least two of the three HS sites accounted for the high transcriptional expression in C2C12 cells. One was located at -2.4 kb and corresponded to a known PEA3/AP1_A element and the other one was located at -4.9 kb and contained a CArG box and a CRE element. So far, no regulatory function had been assigned to this CRE/CArG element. Both HS sites alone were able to activate transcription of a heterologous promoter and showed a cooperative effect when placed together. Electrophoretic mobility-shift assays using myoblast nuclear extracts and specific antibodies demonstrated that cJun, JunD and ATF2 bound to the PEA3/AP1_A element, whereas the CRE/CArG element bound SRF. Altogether, these results suggest that high uPA expression in myoblasts is dependent on the cooperation of two regulatory sites in the uPA promoter.

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