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Allosteric mechanism for site-specific ubiquitination of FANCD2

Viduth K Chaugule1,2, Connor Arkinson3,4, Martin L Rennie3

  • 1Institute of Molecular, Cell and Systems Biology, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, UK. Chaugule.Viduth@gmail.com.

Nature Chemical Biology
|December 25, 2019
PubMed

Insights

Researchers discovered how FANCL activates UBE2T for DNA repair. This mechanism involves unique protein interactions crucial for the Fanconi anemia (FA) pathway, enhancing DNA crosslink repair efficiency.

Area of Science:

  • Molecular Biology
  • Biochemistry
  • Genetics

Background:

  • DNA damage repair is essential for maintaining genomic stability.
  • The Fanconi anemia (FA) pathway repairs DNA interstrand crosslinks via FANCD2/FANCI monoubiquitination.
  • The precise mechanism of ubiquitination by the FA core complex (FA-CC) remains unclear.

Purpose of the Study:

  • To elucidate the mechanism by which FANCL activates UBE2T for FANCD2/FANCI monoubiquitination.
  • To understand how FANCL achieves site-specific ubiquitination within the FA pathway.
  • To explore the potential for engineering UBE2T to enhance ubiquitination activity.

Main Methods:

  • In vitro biochemical assays to study protein-protein interactions and enzymatic activity.
  • Site-directed mutagenesis to probe the roles of specific amino acid residues.
  • Structural analysis to understand the three-dimensional complementarity between UBE2T and FANCD2.

Main Results:

  • FANCL allosterically activates UBE2T, a ubiquitin-conjugating enzyme, to catalyze FANCD2 monoubiquitination.
  • FANCL rewires UBE2T's internal network, altering its active site conformation.
  • A unique basic triad in UBE2T engages an acidic patch on FANCD2, ensuring site-specific ubiquitination.
  • Engineered UBE2T enhances FANCL-mediated di-monoubiquitination of FANCD2/FANCI without losing specificity.

Conclusions:

  • FANCL employs a novel allosteric mechanism to control UBE2T activity for DNA repair.
  • The interaction between UBE2T's active site and FANCD2's surface is critical for site-specific monoubiquitination.
  • Targeted engineering of UBE2T offers a strategy to improve DNA repair pathway efficiency.

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