Identification of glucocorticoid responsive elements (GREs) at far upstream of rat NPY gene

Neurochem Int. 1992 Sep;21(2):185-9. doi: 10.1016/0197-0186(92)90145-h.

Abstract

The location of three glucocorticoid responsive elements (GREs) in rat neuropeptide Y (NPY) gene was determined by chloramphenicol acetyltransferase (CAT) assay and nucleotide sequencing. We have reported that mRNA content of rat prepro-NPY is increased by 1.7-fold in NG108-15 cells by 1 microM dexamethasone, suggesting the presence of GRE in the gene. To identify the element, the 5'-flanking DNA of 3.3 kilobases (kb) was isolated from rat NPY gene. When chimeric chloramphenicol CAT plasmids containing various deletions of the NPY upstream sequence were transfected into NG108-15 cells, the region between -2.9 and -2.1 kb relative to the cap site was found to potentiate the transcription of CAT gene in the presence of 1 microM dexamethasone. The nucleotide sequencing of this region revealed three GRE consensus sequences at -2.5, -2.2 and -2.1 kb. The results indicate that these elements present in the far upstream region of the NPY gene confer induction by glucocorticoids.

MeSH terms

  • Animals
  • Base Sequence
  • Binding Sites
  • Chloramphenicol O-Acetyltransferase / genetics
  • Chloramphenicol O-Acetyltransferase / metabolism
  • Cloning, Molecular
  • DNA / genetics
  • DNA / isolation & purification
  • Dexamethasone / pharmacology*
  • Genes, Regulator / drug effects*
  • Hybrid Cells
  • Mice
  • Molecular Sequence Data
  • Neuropeptide Y / biosynthesis
  • Neuropeptide Y / genetics*
  • Protein Precursors / biosynthesis
  • RNA, Messenger / analysis
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism*
  • Rats
  • Transfection
  • Tumor Cells, Cultured

Substances

  • Neuropeptide Y
  • Protein Precursors
  • RNA, Messenger
  • Dexamethasone
  • DNA
  • preproneuropeptide Y
  • Chloramphenicol O-Acetyltransferase