Identification of nucleolin as a glucocorticoid receptor interacting protein

Biochem Biophys Res Commun. 2001 Jan 19;280(2):476-80. doi: 10.1006/bbrc.2000.4141.

Abstract

The glucocorticoid receptor (GR) is a ligand-induced transcription factor which modulates the transcriptional activity of target genes. Full transcriptional activity of GR is achieved with the help of accessory proteins that are able to interact with GR. We have identified a 95-kDa protein by a blotting technique which utilizes a radioactively labeled DNA-bound GR to detect proteins that bind to this complex. Biochemical purification of this protein followed by protein microsequencing resulted in the identification of human nucleolin. In addition we could show that a GR-deletion mutant localizes to the nucleolus, where nucleolin is one of the most abundant proteins. The binding of nucleolin to this deletion mutant was demonstrated by GST-pull-down experiments. We suggest a biological role of nucleolin in binding of GR in the nucleolus.

MeSH terms

  • Animals
  • COS Cells
  • Cell Nucleolus / chemistry
  • Cell Nucleolus / metabolism
  • HeLa Cells
  • Humans
  • Microscopy, Fluorescence
  • Molecular Weight
  • Nuclear Proteins / chemistry
  • Nuclear Proteins / isolation & purification
  • Nuclear Proteins / metabolism
  • Nucleolin
  • Phosphoproteins / chemistry
  • Phosphoproteins / isolation & purification
  • Phosphoproteins / metabolism*
  • Protein Binding
  • Protein Structure, Tertiary
  • RNA-Binding Proteins / chemistry
  • RNA-Binding Proteins / isolation & purification
  • RNA-Binding Proteins / metabolism*
  • Receptors, Glucocorticoid / chemistry
  • Receptors, Glucocorticoid / genetics
  • Receptors, Glucocorticoid / metabolism*
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / metabolism
  • Sequence Deletion / genetics
  • Transfection

Substances

  • Nuclear Proteins
  • Phosphoproteins
  • RNA-Binding Proteins
  • Receptors, Glucocorticoid
  • Recombinant Fusion Proteins