DUF99 family proteins are novel endonucleases that cleave deoxyuridine on DNA substrates

J Biol Chem. 2024 Nov;300(11):107901. doi: 10.1016/j.jbc.2024.107901. Epub 2024 Oct 18.

Abstract

DNA deamination occurs constantly in a cell and causes DNA damage. As this damage can be deleterious, organisms have evolved many systems to eliminate it, such as Endonuclease V (Endo V). DUF99 family protein contains a domain of unknown function similar to Endo V but has not been experimentally characterized to date. Here, we show that DUF99 family proteins cleave the 3'-side of deoxyuridine (dU) on DNA substrates. Based on phylogenetic analysis, we designated this new protein family as Endonuclease dU (Endo_dU). We also observed that Endo_dU coding gene frequently colocalizes with that of uracil-DNA glycosylase (UDG) in halophilic archaea, and we further performed gene knockout of Endo_dU gene on Haloferax volcanii. The transcription level of UDG gene on Endo_dU knockout strain was increased when induced by sodium bisulfite. Thus, we hypothesize that Endo_dU establishes a new endonuclease family with broad phylogenetic distribution and may participate in DNA repair.

Keywords: DNA deamination; DUF99 protein; Haloferax volcanii; domain of unknown function; endonuclease.

MeSH terms

  • Archaeal Proteins* / chemistry
  • Archaeal Proteins* / genetics
  • Archaeal Proteins* / metabolism
  • DNA / metabolism
  • DNA Repair
  • Deoxyuridine* / metabolism
  • Endonucleases / genetics
  • Endonucleases / metabolism
  • Haloferax volcanii* / genetics
  • Haloferax volcanii* / metabolism
  • Phylogeny*
  • Substrate Specificity
  • Uracil-DNA Glycosidase / genetics
  • Uracil-DNA Glycosidase / metabolism

Substances

  • Archaeal Proteins
  • Deoxyuridine
  • Uracil-DNA Glycosidase
  • Endonucleases
  • DNA