Cloning and characterization of hGMEB1, a novel glucocorticoid modulatory element binding protein

FEBS Lett. 1999 Jun 11;452(3):170-6. doi: 10.1016/s0014-5793(99)00634-1.

Abstract

A 21-bp element called glucocorticoid modulatory element (GME) modulates the glucocorticoid receptor-mediated responses via the binding of an as yet poorly characterized transacting complex of proteins containing the 88-kDa GMEB1 and the 67-kDa GMEB2. Using heat shock protein 27 (HSP27) as bait in the yeast two-hybrid assay, we cloned a 1.83-kb cDNA encoding a novel 573-amino acid protein called human GMEB1 (hGMEB1). hGMEB1 possesses a KDWK domain, contains sequences almost identical (36/38) to three tryptic peptides of rat GMEB1 and shares 38% identity with rat GMEB2. hGMEB1 is ubiquitously expressed as a 85-kDa protein in all cell lines and tissues examined. In vitro translated hGMEB1 bound specifically to GME oligonucleotides yielding a complex of similar size to the complex obtained using rat liver nuclear extracts. Both complexes were supershifted with an antibody specific to hGMEB1. Co-immunoprecipitation experiments confirmed the in vivo interaction of HSP27 with hGMEB1.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3T3 Cells
  • Amino Acid Sequence
  • Animals
  • Binding Sites
  • Cell Line
  • Cloning, Molecular
  • Cricetinae
  • Glutathione Transferase / biosynthesis
  • HSP27 Heat-Shock Proteins
  • HeLa Cells
  • Heat-Shock Proteins*
  • Humans
  • Mice
  • Molecular Chaperones
  • Molecular Sequence Data
  • Neoplasm Proteins / genetics
  • Neoplasm Proteins / metabolism
  • Oligodeoxyribonucleotides / chemistry
  • Oligodeoxyribonucleotides / metabolism
  • Protein Biosynthesis
  • Rats
  • Recombinant Fusion Proteins / biosynthesis
  • Recombinant Fusion Proteins / metabolism
  • Saccharomyces cerevisiae
  • Sequence Alignment
  • Sequence Homology, Amino Acid
  • Transcription Factors / chemistry
  • Transcription Factors / genetics
  • Transcription Factors / metabolism*
  • Transfection

Substances

  • GMEB1 protein, human
  • Gmeb1 protein, mouse
  • HSP27 Heat-Shock Proteins
  • HSPB1 protein, human
  • Heat-Shock Proteins
  • Hsbp1 protein, mouse
  • Hspb1 protein, rat
  • Molecular Chaperones
  • Neoplasm Proteins
  • Oligodeoxyribonucleotides
  • Recombinant Fusion Proteins
  • Transcription Factors
  • Glutathione Transferase

Associated data

  • GENBANK/AF099013