Molecular cloning and characterization of a factor that binds the human glucocorticoid receptor gene and represses its expression

J Biol Chem. 1991 Sep 15;266(26):17333-40.

Abstract

The human DNA binding factor GRF-1, which associates with the promoter region of the glucocorticoid receptor gene (hGR gene), is a repressor of glucocorticoid receptor transcription. The GRF-1 cDNA was cloned using polyclonal antibodies against the purified protein. The deduced amino acid sequence from the cDNA sequences show the presence of three sequence motifs characteristic of a zinc finger and one motif suggestive of a leucine zipper in which 1 cysteine is found instead of all leucines. The GRF-1 expressed in COS-1 cells has a molecular weight of 95,000 and enhances the homologous down-regulation of wild-type hGR gene expression. Biochemical analysis suggests that GRF-1 interaction is sequence specific and that transcriptional efficacy of GRF-1 is regulated through its interaction with specific sequence motif, namely 5'-GAAGGAGGTAGCGAGAAAAGAAACTG-GAGAAACTCGGT.GG-3'. The GRF-1 mRNA is 6.5 kilobases long in rat liver and human MCF-7 cells, and its level is regulated by glucocorticoids.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Cell Line
  • Cloning, Molecular
  • DNA / metabolism
  • DNA-Binding Proteins / genetics*
  • DNA-Binding Proteins / metabolism
  • Deoxyribonuclease I
  • GTP-Binding Proteins*
  • Gene Expression Regulation*
  • Guanine Nucleotide Exchange Factors
  • Humans
  • Molecular Sequence Data
  • Nucleic Acid Hybridization
  • Promoter Regions, Genetic
  • Receptors, Glucocorticoid / genetics*
  • Repressor Proteins / genetics*
  • Restriction Mapping
  • Transcription, Genetic
  • Tumor Suppressor Proteins*
  • ras GTPase-Activating Proteins*
  • ras-GRF1

Substances

  • ARHGAP35 protein, human
  • Arhgap35 protein, rat
  • DNA-Binding Proteins
  • Guanine Nucleotide Exchange Factors
  • RASGRF1 protein, human
  • Receptors, Glucocorticoid
  • Repressor Proteins
  • Tumor Suppressor Proteins
  • ras GTPase-Activating Proteins
  • ras-GRF1
  • DNA
  • Deoxyribonuclease I
  • GTP-Binding Proteins

Associated data

  • GENBANK/M64109
  • GENBANK/M73077
  • GENBANK/S53397
  • GENBANK/S53399
  • GENBANK/S53401
  • GENBANK/S53403
  • GENBANK/S54662
  • GENBANK/S54667
  • GENBANK/S56522
  • GENBANK/S56524