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J. Biol. Chem., Vol. 278, Issue 3, 1533-1541, January 17, 2003

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Characterization of the c-Myb-responsive Region and Regulation of the Human Type I Collagen 2 Chain Gene by c-Myb^*

Michele M. Luchetti^§, Paolo Paroncini, Petra Majlingovà, Jon Frampton^¶, Michael Mucenski^**, Silvia Svegliati Baroni, Paola Sambo, Josee Golay, Martino Introna, and Armando Gabrielli

From the  Laboratorio di Medicina Molecolare, Istituto di Clinica Medica, Ematologia ed Immunologia Clinica, Università di Ancona, 60020 Ancona, Italy, ^¶ Division of Immunity and Infection, Birmingham University Medical School, Birmingham B15 2TT, United Kingdom, ^** Division of Pulmonary Biology, Children's Hospital Medical Center, Cincinnati, Ohio 45229-3039, and  Dipartimento di Immunologia e Biologia Cellulare, Istituto Ricerche Farmacologiche Mario Negri, 20154 Milano, Italy

We have characterized the role of c-Myb and B-Myb in the regulation of human type I collagen 2 chain gene expression in fibroblastic cells. We have identified four Myb-binding sites (MBSs) in the promoter. Transactivation assays on wild type and mutant promoter-reporter constructs demonstrated that c-Myb, but not B-Myb, can transactivate the human type I collagen 2 chain gene promoter via the MBS-containing region. Electrophoretic mobility shift assay experiments showed that c-Myb specifically binds to each of the four MBS; however, the mutagenesis of site MBS-4 completely inhibited transactivation by c-Myb, at least in the full-length promoter. In agreement with these results, c-myb^/ mouse embryo fibroblasts (MEFs) showed a selective lack of expression of type I collagen 2 chain gene but maintained the expression of fibronectin and type III collagen. Furthermore, transforming growth factor- induced type I collagen 2 chain gene expression in c-myb^/ MEFs, implying that the transforming growth factor- signaling pathway is maintained and that the absence of COL1A2 gene expression in c-myb^/ MEFs is a direct consequence of the lack of c-Myb. The demonstration of the importance of c-Myb in the regulation of the type I collagen 2 chain gene suggests that uncontrolled expression of c-Myb could be an underlying mechanism in the pathogenesis of several fibrotic disorders.

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