5' flanking sequence of the human immediate early responsive gene ccn1^* (cyr61) and mapping of polymorphic CA repeat sequence motifs in the human ccn1 (cyr61) locus

N Schütze¹, N Rücker², J Müller¹, J Adamski³ and F Jakob²

¹ Orthopädische Klinik, König-Ludwig-Haus, Labor für Molekulare Experimentelle Orthopädie, Brettreichstrasse 11, 97074 Würzburg, Germany
² Klinische Forschergruppe, Medizinische Poliklinik, Universität Würzburg, Röntgenring 11, 97070 Würzburg, Germany
³ GSF-Forschungszentrum für Umwelt und Gesundheit, Institut für Experimentelle Genetik, Ingolstädter Landstraße 1, 85764 Neuherberg, Germany

Correspondence to:
Dr Schütze n-schuetze.klh{at}mail.uni-wuerzburg.de

Aims—The human ccn1 (hccn; hcyr61) gene has been identified previously at the mRNA and protein level as a 1,25-dihydroxyvitamin D₃ and growth factor regulated gene in human osteoblasts. This study aimed to analyse genomic clones containing the human ccn1 (cyr61) gene and to provide the 5' flanking region.

Methods—Genomic clones were isolated by screening a library and by array filter hybridisations of a genomic library. Sequencing was performed using the dye terminator method. Promoter activity was measured after transient transfection using a ß galactosidase assay. CA repeat motifs were studied by a combined PCR/fragment analysis protocol.

Results—The human 5' flanking region of 870 nucleotides contains several stretches with high homology to the mouse promoter as well as CA repeat motifs. This first report on the human 5' flanking sequence of the hccn1 (hcyr61) gene provides important insights into regulation pathways for the expression of this 1,25-dihydroxyvitamin D₃ and growth factor responsive early gene. A genomic clone containing the hccn1 (hcyr61) gene region also yielded a CA sequence located 3' of the ccn1 (cyr61) gene. This CA repeat and one of the CA repeat motifs in the promoter were studied in detail and found to be polymorphic.

Conclusions—The 5' flanking sequence of the hccn1 (hcyr61) gene provides insights into the mechanisms of regulation of this immediate early gene product. The CA repeat polymorphisms within the gene region will be useful in the genetic study of disorders affecting bone metabolism.

Key Words: human ccn1 (cyr61) gene • gene regulation • promoter • CA repeat polymorphism

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

• You, J.-J., Yang, C.-H., Chen, M.-S., Yang, C.-M. (2009). Cysteine-rich 61, a Member of the CCN Family, as a Factor Involved in the Pathogenesis of Proliferative Diabetic Retinopathy. IOVS 50: 3447-3455 [Abstract] [Full Text]  
• Lee, H.-Y., Chung, J.-W., Youn, S.-W., Kim, J.-Y., Park, K.-W., Koo, B.-K., Oh, B.-H., Park, Y.-B., Chaqour, B., Walsh, K., Kim, H.-S. (2007). Forkhead Transcription Factor FOXO3a Is a Negative Regulator of Angiogenic Immediate Early Gene CYR61, Leading to Inhibition of Vascular Smooth Muscle Cell Proliferation and Neointimal Hyperplasia. Circ. Res. 100: 372-380 [Abstract] [Full Text]  
• Kunz, M., Moeller, S., Koczan, D., Lorenz, P., Wenger, R. H., Glocker, M. O., Thiesen, H.-J., Gross, G., Ibrahim, S. M. (2003). Mechanisms of Hypoxic Gene Regulation of Angiogenesis Factor Cyr61 in Melanoma Cells. J. Biol. Chem. 278: 45651-45660 [Abstract] [Full Text]  
• Liang, Y., Li, C., Guzman, V. M., Evinger, A. J. III, Protzman, C. E., Krauss, A. H.-P., Woodward, D. F. (2003). Comparison of Prostaglandin F2{alpha}, Bimatoprost (Prostamide), and Butaprost (EP2 Agonist) on Cyr61 and Connective Tissue Growth Factor Gene Expression. J. Biol. Chem. 278: 27267-27277 [Abstract] [Full Text]  
• Hilfiker, A., Hilfiker-Kleiner, D., Fuchs, M., Kaminski, K., Lichtenberg, A., Rothkotter, H.-J., Schieffer, B., Drexler, H. (2002). Expression of CYR61, an Angiogenic Immediate Early Gene, in Arteriosclerosis and Its Regulation by Angiotensin II. Circulation 106: 254-260 [Abstract] [Full Text]