Cloning and characterization of hELD/OSA1, a novel BRG1 interacting protein

Biochem J. 2002 May 15;364(Pt 1):255-64. doi: 10.1042/bj3640255.

Abstract

A highly conserved multisubunit enzymic complex, SWI/SNF, participates in the regulation of eukaryote gene expression through its ability to remodel chromatin. While a single component of SWI/SNF, Swi2 or a related protein, can perform this function in vitro, the other components appear to modulate the activity and specificity of the complex in vivo. Here we describe the cloning of hELD/OSA1, a 189 KDa human homologue of Drosophila Eld/Osa protein, a constituent of Drosophila SWI/SNF. By comparing conserved peptide sequences in Eld/Osa homologues we define three domains common to all family members. A putative DNA binding domain, or ARID (AT-rich DNA-interacting domain), may function in targetting SWI/SNF to chromatin. Two other domains unique to Eld/Osa proteins, EHD1 and EHD2, map to the C-terminus. We show that EHD2 mediates binding to Brahma-related gene 1 (BRG1), a human homologue of yeast Swi2. EHD1 and EHD2 also appear capable of interacting with each other. Using an antibody raised against EHD2 of hELD/OSA1, we detected Eld/Osa1 in endogenous SWI/SNF complexes derived from mouse brain.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Motifs
  • Amino Acid Sequence
  • Animals
  • Antibodies, Monoclonal / metabolism
  • Base Sequence
  • COS Cells
  • Cattle
  • Chromatin / metabolism
  • Cloning, Molecular
  • DNA / metabolism
  • DNA Helicases
  • DNA, Complementary / metabolism
  • DNA-Binding Proteins / chemistry*
  • DNA-Binding Proteins / genetics*
  • Drosophila
  • Epitopes
  • Humans
  • Immunoblotting
  • Mice
  • Molecular Sequence Data
  • Nuclear Proteins / chemistry*
  • Nuclear Proteins / genetics*
  • Nuclear Proteins / metabolism*
  • Peptides / chemistry
  • Precipitin Tests
  • Protein Binding
  • Protein Structure, Tertiary
  • Transcription Factors / metabolism*
  • Transfection
  • Two-Hybrid System Techniques

Substances

  • ARID1B protein, human
  • Antibodies, Monoclonal
  • Arid1a protein, mouse
  • Chromatin
  • DNA, Complementary
  • DNA-Binding Proteins
  • Epitopes
  • Nuclear Proteins
  • Peptides
  • Transcription Factors
  • DNA
  • SMARCA4 protein, human
  • Smarca4 protein, mouse
  • DNA Helicases