Beta-adrenergic stimulation of cFOS via protein kinase A is mediated by cAMP regulatory element binding protein (CREB)-dependent and tissue-specific CREB-independent mechanisms in corticotrope cells

J Biol Chem. 1992 Nov 25;267(33):23520-6.

Abstract

Catecholamines stimulate proopiomelanocortin (POMC) gene expression in corticotrope cells, but the molecular mechanisms of these effects are not known. While beta-adrenergic receptors stimulate the protein kinase A (PKA) system, the POMC promoter does not have classical cAMP-response elements (CREs). Therefore, we investigated the induction of the c-fos protooncogen, previously shown to increase POMC transcription in AtT20 cells. In this corticotrope-derived cell line, we show that activation of beta-receptors with isoprenaline (Iso) induces a transient rise in c-fos mRNA levels. Gel mobility shift assays with a labeled AP1 consensus sequence (TGACTCA) showed induction of specific binding activity after Iso treatment. Cotransfection experiments with dominant inhibitory PKA mutants and reporter genes containing c-fos promoter sequences showed that c-fos induction by Iso is entirely dependent on a functional PKA activity. Furthermore, we show that beta-receptor induction of c-fos in corticotrophs is mediated by at least two distinct cAMP-responsive sequences. cAMP regulatory element binding (CREB)-dependent induction is observed on the CRE located at -60 bp on the c-fos promoter. A region located in the vicinity of the dyad symetry element (-290) is also found to mediate tissue-specific cAMP induction. Transcriptional activation by this site, although sensitive to PKA antagonism, is not blocked by CREB mutants.

Publication types

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

MeSH terms

  • Animals
  • Base Sequence
  • Cell Nucleus / metabolism
  • Chloramphenicol O-Acetyltransferase / genetics
  • Chloramphenicol O-Acetyltransferase / metabolism
  • Cyclic AMP / physiology
  • Cyclic AMP Response Element-Binding Protein / metabolism*
  • Gene Expression Regulation, Neoplastic
  • Genes, fos*
  • Humans
  • Isoproterenol / pharmacology*
  • Kinetics
  • Metallothionein / genetics
  • Mice
  • Molecular Sequence Data
  • Oligodeoxyribonucleotides
  • Pituitary Neoplasms
  • Pro-Opiomelanocortin / genetics*
  • Promoter Regions, Genetic*
  • Protein Kinases / metabolism*
  • Proto-Oncogene Proteins c-fos / biosynthesis
  • Proto-Oncogene Proteins c-fos / genetics*
  • RNA, Messenger / metabolism
  • Receptors, Adrenergic, beta / physiology*
  • Sequence Deletion
  • Transfection
  • Tumor Cells, Cultured

Substances

  • Cyclic AMP Response Element-Binding Protein
  • Oligodeoxyribonucleotides
  • Proto-Oncogene Proteins c-fos
  • RNA, Messenger
  • Receptors, Adrenergic, beta
  • Pro-Opiomelanocortin
  • Metallothionein
  • Cyclic AMP
  • Chloramphenicol O-Acetyltransferase
  • Protein Kinases
  • Isoproterenol