THE JOI~NAL OF BIOLOGICAL CHEMISTRY Vol. 269, No. 14, Issue of April 8, pp. 10363-10369, 1994 0 1994 by The American Society for Biochemistry and Molecular Biology, Inc. Printed in U.S.A. Phenotypic Alterations in Fos-transgenic Mice Correlate with Changes in Fos/Jun-dependent Collagenase Type I Expression REGULATION OF MOUSE METALLOPROTEINASES BY CARCINOGENS, TUMOR PROMOTERS, CAMP, AND FOS ONCOPROTEIN* (Received for publication, December 20, 1993) Sabine GackS, Riidiger VallonS, Jorg SchaperS, Ulrich RiitherQ, and Peter Angelh From the $Kernforschungszentrum Karlsruhe, Znstitut fur Genetik, Postfach 3640, 76021 Karlsruhe, Federal Republic of Germany and the PMedizinische Hochschule Hannouer, Znstitut fur Molekularbiologie, 30623 Hannover, Federal Republic of Germany Using specific cDNAs isolated from mouse fibroblasts we determined tissue-specific expression of different matrix metalloproteinase genes: both stromelysin-1 and collagenase IV are highly expressed in heart and lung, whereas collagenase I is expressed most abundantly in skeletal muscle, kidney, and bone. High basal level ex- pression of stromelysin-2 is found in heart and kidney. Like in man and rat, the expressions of collagenase I, stromelysin-1, and stromelysin-2 are regulated by the tumor promoter 12-0-tetradecanoyl-phorbol 13-acetate and by W irradiation, but not by cAMF? In contrast, the expression of the 72-kDa collagenase IV is not affected by either stimuli. We and others have shown previously that under cell culture conditions, the regulation of hu- man collagenase I is regulated by the transcription fac- tor FodJun (AP-1). Here we show that in c-foetransgenic mice transcription of collagenase I is induced in thymus, spleen, and, most dominantly,in bone upon overexpres- sion of Fos. Neither collagenase IV nor stromelysin-1 or stromelysin-2 expression is affected by c-Fos. The sites of induced collagenase I expression correlate with the sites of Fos-induced long-term cellular alterations in transgenic mice including bone remodeling and T cell development. In fact, in the developing bone tumors strongly enhanced levels of collagenase I transcripts were detectable. These results identify collagenase I as a Fos-regulated gene in vivo and suggest a possible role for FodJun heterodimers in establishing the pathologi- cal phenotypeof c-fos transgenic mice. Resident cells of tissues are capable of secreting an array of structurally related zinc endopeptidases known as matrix me- talloproteinases (MMPs).’ The MMPs initiate the degradation of the surrounding macromolecules of the extracellular matrix (ECM), mostly proteoglycans and the different specific types of collagen. ECM degradation presumably contributes to the ini- tial phase of tissue remodeling inherent to the physiological processes of morphogenesis,angiogenesis, involution of the uterus, bone resorption, inflammation, andwound healing (for * This work was supported by a grant from the Deutsche Forsch- ungsgemeinschaft (An 182/6-1). The costs of publication of this article were defrayed in part by the payment of page charges. This article must U.S.C. Section 1734 solely to indicate this fact. therefore be hereby marked “advertisement” in accordance with 18 ll To whom correspondence should be addressed: Kernforschungszen- trum Karlsruhe, Institut fir Genetik, Postfach 3640, 76021 Karlsruhe, Germany, Tel.: 07247-823-444; Fax: 07247-823-354. The abbreviations used are: MMP, matrix metalloproteinase; ECM, extracellular matrix; TPA, 12-0-tetradecanoylphorbol-13-acetate; PCR, polymerase chain reaction; kb, kilobase paids); TIMP, tissue inhibitor of metalloproteinases. review, see Refs. 1-5). Aberrant activity of MMPs has been found to be involved in a variety of pathological processes such as rheumatoid joint destruction, corneal ulceration, metastasis of tumor cells, and genetic diseases (e.g. recessive dystrophic epidermolysis bullosa; for review, see Refs. 3-6). The various members of the MMP family contain several distinct domains that are highly conserved. Despite this high degree of similarity, the MMPs identified so far differ in sub- strate specificity and expression in response to extracellular stimuli, suggesting that each individual member of the MMP family has a distinct function in the physiological and patho- logical processes listed above (1-3, 6). Furthermore, the activ- ity of these enzymes is tightly regulated on several levels: regu- lation of transcription (7-151, activation of the latent pro- enzyme (16-191, and interaction with specific inhibitors of MMPs (TIMP-1 and TIMP-2, Refs. 20-22). Studies in cell cul- ture systems have revealed multiple mechanisms that posi- tively or negatively interfere with the transcription of MMPs. Transcription is enhanced by carcinogens, cytokines, and tu- mor promoters (for review, see Refs. 4 and 23); and repressed by steroid hormones (24-26) and by the E1A products of adenovi- rus (25, 27, 28). The transcription of human collagenase I and of human and rat stromelysin-1 are regulated by the transcrip- tion factor AP-1 in most of the cell systems examined so far (10, 29-32). AP-1 is a heterodimeric complex whose subunits are encoded by members of the jun, fos, and ATF gene families (for review, see Ref. 23). AP-1 activity is repressed, presumably by physical interaction with the activated steroid hormone recep- tors (26, 33). Despite the great body of information of MMP expression in cultured cells, little is known about the expression and function of distinct MMPs in the intact multicellular organism. For ex- ample, expression of collagenase type I has been found to be regulated by basic fibroblast growth factor and transforming growth factor p (12, 341, both of which play a decisive role during early development (35). In fact, changes in the expres- sion and activity of MMPs have been observed during in vitro differentiation of mouse F9 and PSA-1 cells, and during peri- implantation development of mouse embryos (36). Spatial and cell type-specific expression in adult mice or during mouse de- velopment (e.g. by in situ hybridization) has been examined only for the 72-kDa form of collagenase type IV (37). To be able to dissect the complex regulation of MMP gene expression in mouse embryos and adult mice, and to determine the mechanisms of transcriptional regulation of mouse MMP expression, we isolated specific mouse probes encoding MMP-1 (collagenase type I), MMP-2 (72-kDa collagenase type IV), MMP-3 (stromelysin-l), and MMP-10 (stromelysin-2).These clones could be unequivocally assigned by their homologies to rat and man. High degrees of tissue-specific expression support 10363