Oligonucleotide squelching reveals the mechanism of estrogen receptor autologous down-regulation

Mol Endocrinol. 1997 Jun;11(7):938-49. doi: 10.1210/mend.11.7.9936.

Abstract

Antisense oligos complementary to the 5'-end, but not to the 3'-end, of the estrogen receptor (ER) messenger RNA caused a paradox accumulation of ER protein in MCF-7 cells. The same effect was observed after treatment of the cells with the corresponding sense oligos. The oligos interfering with ER down-regulation were demonstrated to specifically bind the ER with affinities in the nanomolar range. It is, therefore, proposed that the ER up-regulation induced by the oligos might be due to squelching of the ER (or ER-inducible proteins) from their binding site located in the 5'-end of the ER gene. We also report that transcriptionally inactive ER mutants can undergo down-regulation, and that in denaturing gels, the migration profile of ER-oligo and ER-estrogen-responsive element complexes are dissimilar. We, therefore, propose that ER can interact with DNA in different ways and at different binding sites. These observations might have important pharmacological consequences, since specific drugs could be devised to induce the ER conformation necessary to perform only selected tasks of the ER transcriptional repertoire.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Blotting, Western
  • COS Cells
  • DNA Primers / chemistry
  • Down-Regulation / genetics*
  • Electrophoresis, Polyacrylamide Gel
  • Gene Deletion
  • Humans
  • Luciferases / genetics
  • Oligonucleotides / chemistry
  • Oligonucleotides / genetics
  • Oligonucleotides / pharmacology
  • Oligonucleotides, Antisense / chemistry
  • Oligonucleotides, Antisense / genetics*
  • Oligonucleotides, Antisense / pharmacology
  • Phosphorus Radioisotopes
  • Receptors, Estrogen / biosynthesis*
  • Receptors, Estrogen / genetics
  • Receptors, Estrogen / metabolism
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Transcription, Genetic / physiology
  • Transfection / genetics*
  • Tumor Cells, Cultured
  • Urea / chemistry

Substances

  • DNA Primers
  • Oligonucleotides
  • Oligonucleotides, Antisense
  • Phosphorus Radioisotopes
  • Receptors, Estrogen
  • Recombinant Proteins
  • Urea
  • Luciferases