Conserved function of Drosophila Fancd2 monoubiquitination in response to double-strand DNA breaks

G3 (Bethesda). 2022 Jul 29;12(8):jkac129. doi: 10.1093/g3journal/jkac129.


Fanconi anemia genes play key roles in metazoan DNA damage responses, and human FA mutations cause numerous disease phenotypes. In human cells, activating monoubiquitination of the Fanconi anemia protein Fancd2 occurs following diverse DNA damage stimuli. Monoubiquitinated Fancd2 forms nuclear foci to recruit additional repair factors. Fancd2 animal models to date have focused on molecular nulls or whole gene knockdown, leaving the specific in vivo role of monoubiquitination unclear. Using a point mutant in a conserved residue, we recently linked Drosophila Fancd2 monoubiquitination to a mitosis-specific DNA double-strand break response. In this context, we used CRISPR/Cas9 to generate the first animal model of an endogenous mutation in the conserved monoubiquitination site (fancd2K595R). Here, we expand upon our characterization of fancd2K595R. We also introduce and characterize additional Drosophila tools to study fancd2, including new mutant alleles and GFP-tagged rescue transgenes. Using these new reagents, we show the impact of Drosophila Fancd2 on organismal and cell viability, as well as on repair protein localization, in the presence or absence of double-strand breaks. These findings expand our understanding of Fanconi anemia gene function in vivo and provide useful reagents for DNA repair research.

Keywords: Drosophila; Fancd2; Mre11; double-strand break; monoubiquitination; radiation.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • DNA Breaks, Double-Stranded*
  • DNA Damage
  • DNA Repair / genetics
  • Drosophila Proteins* / genetics
  • Drosophila Proteins* / metabolism
  • Drosophila* / genetics
  • Drosophila* / metabolism
  • Fanconi Anemia / genetics
  • Fanconi Anemia / metabolism
  • Fanconi Anemia Complementation Group D2 Protein* / genetics
  • Fanconi Anemia Complementation Group D2 Protein* / metabolism
  • Humans
  • Ubiquitination


  • Drosophila Proteins
  • FANCD2 protein, human
  • Fancd2 protein, Drosophila
  • Fanconi Anemia Complementation Group D2 Protein