Murine Matrix Metalloproteinase 9 Gene

doi:10.1074/jbc.274.9.5588

Murine Matrix Metalloproteinase 9 Gene

5′-UPSTREAM REGION CONTAINS CIS-ACTING ELEMENTS FOR EXPRESSION IN OSTEOCLASTS AND MIGRATING KERATINOCYTES IN TRANSGENIC MICE*

From the ^‡Department of Biology, University of Liege, B-4000 Liege, Belgium, ^¶Biocenter and Department of Biochemistry, University of Oulu, FIN-90570 Oulu, Finland, and the ^‖Division of Matrix Biology, Department of Medical Biochemistry and Biophysics, Karolinska Institute, S-171 77 Stockholm, Sweden

Abstract

Knowledge about the regulation of cell lineage-specific expression of extracellular matrix metalloproteinases is limited. In the present work, the murine matrix metalloproteinase 9 (MMP-9) gene was shown to contain 13 exons, and the 2.8-kilobase pair upstream region was found to contain several common promoter elements including a TATA box-like motif, three GC boxes, four AP-1-like binding sites, an AP-2 site, and three PEA3 consensus sequences that may be important for basic activity of the gene. In order to identify cell-specific regulatory elements, constructs containing varying lengths of the upstream region in front of a LacZ reporter gene were made and studied for expression in transgenic mice generated by microinjection into fertilized oocytes. Analyses of the mice revealed that the presence of sequences between −2722 and −7745 allowed for expression in osteoclasts and migrating keratinocytes,i.e. cells that have been shown to normally express the enzyme in vivo. The results represent the first in vivo demonstration of the location of cell-specific control elements in a matrix metalloproteinase gene and show that element(s) regulating most cell-specific activities of 92-kDa type collagenase are located in the −2722 to −7745 base pair region.

Footnotes

↵* This work was supported in part by grants from the Academy of Finland, the Swedish Medical Research Council, the Swedish Cancer Society, Hedlundı́s Foundation, EU Biomed II Grant BMH4-CT96–0017, the Fonds de la Recherche Scientifique Médicale, the Fonds National de la Recherche Scientifique, the Center Anticancéreux prés l’Université de Liège, and the CGER-Assurances 1996/1999.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
↵§ These two authors contributed equally to this work.
↵** To whom correspondence should be addressed: Division of Matrix Biology, Dept. of Medical Biochemistry and Biophysics, Karolinska Institute, S-171 77 Stockholm, Sweden. Tel.: 46-8-728-7720; Fax: 46-8-31-61-65; E-mail: karl.tryggvason{at}mbb.ki.se.
↵2 C. Munaut, T. Salonurmi, S. Kontusaari, P. Reponen, T. Morita, J.-M. Foidart, and K. Tryggvason, unpublished results.
Abbreviations:

MMP

matrix metalloproteinase

X-gal

5-bromo-4-chloro-3-indolyl-β-galactopyronoside

kb

kilobase pair(s)

bp

base pair(s)
- Received May 6, 1998.
- Revision received July 23, 1998.
The American Society for Biochemistry and Molecular Biology, Inc.