Identification of NGF-response element in the rat neuropeptide Y gene and induction of the binding proteins

Biochem Biophys Res Commun. 1992 Dec 30;189(3):1553-60. doi: 10.1016/0006-291x(92)90253-h.

Abstract

Gene expression of the rat neuropeptide Y (NPY) increases by 100 times, as the PC12 cells differentiate into sympathetic neuron-like cells with NGF treatment and this increase is partly due to transcriptional activation of the NPY gene (Sabol and Higuchi, Mol. Endocrinol. 4, 384, 1990). To identify the NGF-response element, a transient expression assay was carried out by using the CAT reporter genes containing various lengths of the 5' upstream region of the NPY gene in the PC12 cells. The 48-base element (-80/-33 upstream of the Cap site) was identified as a NGF-response element (NGFRE). Gel shift assay indicated the existence of at least two DNA-binding proteins to NGFRE. The binding activity of the protein(s) (NDF1) to the upper region (-80/-63) was increased by 3-fold with NGF treatment for 24 h. These findings suggest that these nuclear proteins are involved in the enhanced transcription of the NPY gene by NGF.

MeSH terms

  • Animals
  • Base Sequence
  • Binding Sites
  • Cell Differentiation / drug effects
  • Chloramphenicol O-Acetyltransferase / genetics
  • Chloramphenicol O-Acetyltransferase / metabolism
  • Cloning, Molecular
  • Molecular Sequence Data
  • Nerve Growth Factors / pharmacology*
  • Neuropeptide Y / genetics*
  • Oligodeoxyribonucleotides / isolation & purification
  • PC12 Cells
  • Plasmids
  • Promoter Regions, Genetic*
  • Rats
  • Receptors, Neuropeptide Y / biosynthesis*
  • Recombinant Proteins / metabolism
  • Restriction Mapping
  • Sequence Deletion
  • Transcription, Genetic / drug effects

Substances

  • Nerve Growth Factors
  • Neuropeptide Y
  • Oligodeoxyribonucleotides
  • Receptors, Neuropeptide Y
  • Recombinant Proteins
  • Chloramphenicol O-Acetyltransferase