Cellular repair of DNA–DNA cross-links induced by 1,2,3,4-Diepoxybutane

Lisa N. Chesner, Amanda Degner, Dewakar Sangaraju, Shira Yomtoubian, Susith Wickramaratne, Bhaskar Malayappan, Natalia Tretyakova, Colin Campbell

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5 Scopus citations

Abstract

Xenobiotic-induced interstrand DNA–DNA cross-links (ICL) interfere with transcription and replication and can be converted to toxic DNA double strand breaks. In this work, we investigated cellular responses to 1,4-bis-(guan-7-yl)-2,3-butanediol (bis-N7G-BD) cross-links induced by 1,2,3,4-diepoxybutane (DEB). High pressure liquid chromatography electrospray ionization tandem mass spectrometry (HPLC-ESI+-MS/MS) assays were used to quantify the formation and repair of bis-N7G-BD cross-links in wild-type Chinese hamster lung fibroblasts (V79) and the corresponding isogenic clones V-H1 and V-H4, deficient in the XPD and FANCA genes, respectively. Both V-H1 and V-H4 cells exhibited enhanced sensitivity to DEB-induced cell death and elevated bis-N7G-BD cross-links. However, relatively modest increases of bis-N7G-BD adduct levels in V-H4 clones did not correlate with their hypersensitivity to DEB. Further, bis-N7G-BD levels were not elevated in DEB-treated human clones with defects in the XPA or FANCD2 genes. Comet assays and γ-H2AX focus analyses conducted with hamster cells revealed that ICL removal was associated with chromosomal double strand break formation, and that these breaks persisted in V-H4 cells as compared to control cells. Our findings suggest that ICL repair in cells with defects in the Fanconi anemia repair pathway is associated with aberrant re-joining of repair-induced double strand breaks, potentially resulting in lethal chromosome rearrangements.

Original languageEnglish (US)
Article number1086
JournalInternational journal of molecular sciences
Volume18
Issue number5
DOIs
StatePublished - May 18 2017

Bibliographical note

Funding Information:
This work was funded by the National Institutes of Health (CA100670 to N.T., ES023350 to Natalia Tretyakova and Colin Campbell). Lisa N. Chesner is supported by Training Grant 5T32HL007741.

Publisher Copyright:
© 2017 by the authors. Licensee MDPI, Basel, Switzerland.

Keywords

  • 1,2,3,4-diepoxybutane
  • Chinese hamster lung fibroblast
  • DNA double strand break
  • DNA repair
  • Fanconi anemia
  • Homologous recombination
  • Interstrand DNA-DNA crosslink
  • Nucleotide excision repair

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