Structure and hemimethylated CpG binding of the SRA domain from human UHRF1

J Biol Chem. 2008 Dec 12;283(50):34490-4. doi: 10.1074/jbc.C800169200. Epub 2008 Oct 22.

Abstract

Human UHRF1 (ubiquitin-like PHD and RING finger 1) functions to maintain CpG DNA methylation patterns through DNA replication by co-localizing with the DNA methyltransferase DNMT1 at chromatin in mammals. Recent studies show that UHRF1 binds selectively to hemimethylated CpG via its conserved SRA (SET- and RING finger-associated) domain. However, the underlying molecular mechanism is not known. Here, we report a 1.95 A resolution crystal structure of the SRA domain of human UHRF1. Using NMR structure-guided mutagenesis, electrophoretic mobility shift assay, and fluorescence anisotropy analysis, we determined key amino acid residues for methyl-DNA binding that are conserved in the SRA domain.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Anisotropy
  • CCAAT-Enhancer-Binding Proteins / genetics*
  • CCAAT-Enhancer-Binding Proteins / physiology*
  • CpG Islands*
  • Crystallography, X-Ray / methods
  • DNA / chemistry
  • Escherichia coli / metabolism
  • Humans
  • Magnetic Resonance Spectroscopy
  • Models, Molecular
  • Molecular Conformation
  • Mutagenesis
  • Protein Binding
  • Protein Structure, Tertiary
  • Spectrometry, Fluorescence / methods
  • Temperature
  • Ubiquitin-Protein Ligases

Substances

  • CCAAT-Enhancer-Binding Proteins
  • DNA
  • UHRF1 protein, human
  • Ubiquitin-Protein Ligases

Associated data

  • PDB/3DWH