The isolated C-terminal domain of Ring1B is a dimer made of stable, well-structured monomers

Biochemistry. 2007 Nov 6;46(44):12764-76. doi: 10.1021/bi701343q. Epub 2007 Oct 13.

Abstract

The Ring1B is a core subunit protein of the PRC1 (polycomb repressive complex 1), which plays key roles in the regulation of the Homeobox gene expression, X-chromosome inactivation, stem cell self-renewal, and tumorigenesis. The C-terminal region of Ring1B interacts with RYBP, a transcriptional repressor in transiently transfected cells, and also with M33, another transcriptional repressor involved in mesoderm patterning. In this work, we show that the C-terminal domain of Ring1B, C-Ring1B, is a dimer in solution, with a dissociation constant of 200 microM, as shown by NMR, ITC, and analytical gel filtration. Each monomer is stable at physiological conditions in a wide pH range ( approximately 5 kcal mol-1 at 298 K), with a well-formed core and a spherical shape. The dimer has a high content of alpha-helix and beta-sheet, as indicated by FTIR spectra, and it is formed by the mutual docking of the preformed folded monomers. Since the C-terminal region is important for interaction with other proteins of the PRC1, the dimerization and the presence of those well-structured monomers might be a form of regulation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Chromatography, Gel
  • DNA-Binding Proteins / chemistry*
  • DNA-Binding Proteins / isolation & purification
  • DNA-Binding Proteins / metabolism*
  • Dimerization
  • Hydrogen-Ion Concentration
  • Mice
  • Molecular Sequence Data
  • Polycomb Repressive Complex 1
  • Protein Binding
  • Protein Denaturation
  • Protein Folding
  • Protein Structure, Tertiary
  • Repressor Proteins
  • Ubiquitin-Protein Ligases

Substances

  • DNA-Binding Proteins
  • Repressor Proteins
  • Polycomb Repressive Complex 1
  • Rnf2 protein, mouse
  • Ubiquitin-Protein Ligases