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The identification of FANCD2 DNA binding domains reveals nuclear localization sequences

Joshi Niraj1,2, Marie-Christine Caron1,2, Karine Drapeau1,2

  • 1Genome Stability Laboratory, CHU de Québec Research Center, HDQ Pavilion, Oncology Axis, 9 McMahon, Québec City, QC G1R 2J6, Canada.

Nucleic Acids Research
|July 2, 2017
PubMed

Insights

Fanconi anemia protein FANCD2 has key DNA binding domains and nuclear localization sequences. These regions are crucial for FANCD2 function in DNA repair and Fanconi anemia pathway activation.

Area of Science:

  • Genetics
  • Molecular Biology
  • Biochemistry

Background:

  • Fanconi anemia (FA) is a genetic disorder causing bone-marrow failure and cancer susceptibility.
  • The FA pathway involves over 20 FANC genes, crucial for DNA interstrand crosslink resistance.
  • FANCD2 is central to the FA pathway, accumulating on chromatin after DNA damage.

Purpose of the Study:

  • To identify and characterize the DNA binding regions of FANCD2.
  • To investigate the role of nuclear localization sequences (NLS) within FANCD2.
  • To understand how these domains contribute to the FA pathway's efficiency.

Main Methods:

  • Synthetic peptide mapping was used to analyze FANCD2.
  • Electrophoretic mobility shift assays (EMSA) screened for DNA binding domains.
  • Mutagenesis studies assessed the impact of mutations in DNA binding/NLS domains.

Main Results:

  • FANCD2 possesses two major DNA binding domains rich in conserved lysine residues.
  • One N-terminal domain contains both DNA binding and nuclear localization sequences.
  • Mutations in this bifunctional domain impaired FANCD2 monoubiquitination and increased sensitivity to mitomycin C.

Conclusions:

  • DNA binding domains and NLS are critical for FANCD2 function in the Fanconi anemia pathway.
  • The interplay between DNA binding and nuclear import is essential for efficient DNA repair.
  • These findings elucidate key mechanisms underlying Fanconi anemia pathogenesis.

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