Parathyroid Hormone Regulates the Rat Collagenase-3 Promoter in Osteoblastic Cells through the Cooperative Interaction of the Activator Protein-1 Site and the runt Domain Binding Sequence*
- Nagarajan Selvamurugan,
- Wan-Yin Chou‡,
- A. Terrece Pearman§,
- Malini R. Pulumati and
- Nicola C. Partridge¶
- From the Department of Pharmacological and Physiological Science, Saint Louis University School of Medicine, St. Louis, Missouri 63104
Abstract
Parathyroid hormone induces collagenase-3 gene transcription in rat osteoblastic cells. Here, we characterized the basal, parathyroid hormone regulatory regions of the rat collagenase-3 gene and the proteins involved in this regulation. The minimal parathyroid hormone-responsive region was observed to be between base pairs −38 and −148. Deleted and mutated constructs showed that the activator protein-1 and the runt domain binding sites are both required for basal expression and parathyroid hormone activation of this gene. The runt domain site is identical to an osteoblast-specific element-2 or acute myelogenous leukemia binding sequence in the mouse and rat osteocalcin genes, respectively. Overexpression of an acute myelogenous leukemia-1 repressor protein inhibited parathyroid hormone activation of the promoter, indicating a requirement of acute myelogenous leukemia-related factor(s) for this activity. Overexpression of c-Fos, c-Jun, osteoblast-specific factor-2, and core binding factor-β increased the response to parathyroid hormone of the wild type (−148) promoter but not with mutation of either or both the activator protein-1 and runt domain binding sites. In summary, we conclude that there is a cooperative interaction of acute myelogenous leukemia/polyomavirus enhancer-binding protein-2-related factor(s) binding to the runt domain binding site with members of the activator protein-1 transcription factor family binding to the activator protein-1 site in the rat collagenase-3 gene in response to parathyroid hormone in osteoblastic cells.
Footnotes
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↵* This work was supported by National Institutes of Health Grants DK47420 and DK48109 (to N. C. P).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.
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↵‡ Present address: Lilly Research Laboratories, Lilly Corporate Center, Indianapolis, IN 46285.
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↵§ Present address: Human Molecular Biology and Genetics, Eccles Institute of Human Genetics, University of Utah, Salt Lake City, UT 84112.
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↵¶ To whom correspondence should be addressed: Dept. of Pharmacological and Physiological Science, Saint Louis University School of Medicine, 1402 S. Grand Blvd., St. Louis, MO 63104. Tel.: 314-577-8239; Fax: 314-577-8233; E-mail: Partrinc@SLU.EDU.
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↵1 The abbreviations used are: PTH, parathyroid hormone; C/EBP, CCAAT enhancer-binding protein; RD, runtdomain; PEA-3, polyomavirus enhancer activator-3; AP, activator protein; AML-1, acute myelogenous leukemia-1; PEBP, polyomavirus enhancer-binding protein; OSF2, osteoblast-specific factor-2; CREB, cyclic AMP-response element-binding protein; CBF, core binding factor; CAT, chloramphenicol acetyltransferase; ATF-1, activating transcription factor-1; PCR, polymerase chain reaction; MMP, matrix metalloproteinase; ETO, eight-twenty-one.
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↵2 N. Selvamurugan, M. R. Pulumati, and N. C. Partridge, unpublished observations.
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- Received March 25, 1997.
- Revision received January 8, 1998.
- The American Society for Biochemistry and Molecular Biology, Inc.
