Chemically Ubiquitylated PCNA as a Probe for Eukaryotic Translesion DNA Synthesis

Nat Chem Biol. 2010 Apr;6(4):270-2. doi: 10.1038/nchembio.316. Epub 2010 Mar 7.

Abstract

The rapid growth in ubiquitin biology requires facile chemical approaches for protein ubiquitylation that can overcome the common problem of low yield faced by the enzymatic reaction catalyzed by ubiquitin ligases. We report a chemical approach for monoubiquitylation and SUMOylation of PCNA through disulfide exchange and intein chemistry. We used the chemically ubiquitylated and SUMOylated PCNAs in studying translesion DNA synthesis and revealed a surprising degree of flexibility of the ubiquitin modification.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cysteine / metabolism
  • DNA / biosynthesis*
  • DNA / chemistry
  • DNA-Directed DNA Polymerase / metabolism
  • Disulfides / chemistry
  • Disulfides / metabolism
  • Eukaryotic Cells / metabolism*
  • Inteins
  • Molecular Probes / chemistry
  • Molecular Probes / metabolism*
  • Proliferating Cell Nuclear Antigen / chemistry*
  • Proliferating Cell Nuclear Antigen / genetics
  • Proliferating Cell Nuclear Antigen / metabolism*
  • Protein Binding
  • Ubiquitin / metabolism*
  • Ubiquitin-Protein Ligases / metabolism
  • Ubiquitinated Proteins / chemistry
  • Ubiquitinated Proteins / metabolism*
  • Ubiquitination

Substances

  • Disulfides
  • Molecular Probes
  • Proliferating Cell Nuclear Antigen
  • Ubiquitin
  • Ubiquitinated Proteins
  • DNA
  • Ubiquitin-Protein Ligases
  • DNA-Directed DNA Polymerase
  • Cysteine

Associated data

  • PubChem-Substance/87464908
  • PubChem-Substance/87464909