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

J Biol Chem. 2002 Dec 27;277(52):50535-42. doi: 10.1074/jbc.M210526200. Epub 2002 Oct 28.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Base Sequence
  • Calcium Channels, L-Type / drug effects
  • Calcium Channels, L-Type / genetics*
  • Cloning, Molecular
  • Cyclic AMP Response Element-Binding Protein / metabolism*
  • Gene Expression Regulation / drug effects
  • Genes, Reporter
  • Insulin-Like Growth Factor I / pharmacology*
  • Luciferases / genetics
  • Mice
  • Molecular Sequence Data
  • Muscle, Skeletal / physiology*
  • Oligodeoxyribonucleotides, Antisense / pharmacology
  • Promoter Regions, Genetic
  • RNA, Messenger / genetics
  • Transcription, Genetic / drug effects*

Substances

  • Calcium Channels, L-Type
  • Cyclic AMP Response Element-Binding Protein
  • Oligodeoxyribonucleotides, Antisense
  • RNA, Messenger
  • Insulin-Like Growth Factor I
  • Luciferases

Associated data

  • GENBANK/AF343753