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Originally published In Press as doi:10.1074/jbc.M210526200 on October 28, 2002

J. Biol. Chem., Vol. 277, Issue 52, 50535-50542, December 27, 2002
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Insulin-like Growth Factor-1 Increases Skeletal Muscle Dihydropyridine Receptor alpha 1S Transcriptional Activity by Acting on the cAMP-response Element-binding Protein Element of the Promoter Region*

Zhenlin ZhengDagger , Zhong-Min WangDagger , and Osvaldo DelbonoDagger §||

From the Dagger  Department of Physiology and Pharmacology, § Department of Internal Medicine, Gerontology, and  Neuroscience Program, Wake Forest University School of Medicine, Winston-Salem, North Carolina 27157

Previous work from our laboratory has shown that insulin-like growth factor 1 (IGF-1) increases the expression of the skeletal muscle dihydropyridine receptor (DHPR) alpha 1 subunit by regulating DHPR alpha 1S nuclear transcription. In this study, we investigated the mechanism by which IGF-1 enhances expression of the DHPR alpha 1S gene. To this end, the promoter region of the mouse DHPR alpha 1S gene was recently cloned and sequenced and various promoter deletion-luciferase reporter constructs were used. These constructs were transfected into C2C12 cells and IGF-1 effects were measured by recording luciferase activity. IGF-1 significantly enhanced DHPR alpha 1S transcription in those constructs carrying cAMP-response element-binding protein (CREB) binding site but not in CREB core binding site mutants. Gel mobility shift assay using a double stranded oligonucleotide for the CREB site in the promoter region, and competition experiments with excess unlabeled or mutated promoter oligonucleotide, and unlabeled consensus CREB oligonucleotide demonstrated that IGF-1 induces CREB binding to the DHPR alpha 1S promoter. IGF-1-mediated enhancement in charge movement was prevented by incubating the cells with antisense but not with sense oligonucleotides against CREB. These results support the conclusion that IGF-1 regulates DHPR alpha 1S transcription in muscle cells by acting on the CREB element of the promoter.

* This work was supported by NIA National Institutes of Health Grants AG18755, AG13934, and AG15820, and a grant from the Muscular Dystrophy Association of America (to O. D.).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.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EBI Data Bank with accession number(s) AF343753.

|| To whom correspondence should be addressed: Dept. of Physiology and Pharmacology, Wake Forest University School of Medicine, Medical Center Blvd., Winston-Salem, NC 27157. Tel.: 336-716-9802; Fax: 336-716-7359; E-mail: odelbono@wfubmc.edu.

Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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