Cip29 is phosphorylated following activation of the DNA damage response in Xenopus egg extracts

PLoS One. 2017 Jul 17;12(7):e0181131. doi: 10.1371/journal.pone.0181131. eCollection 2017.

Abstract

Acting through a complex signalling network, DNA lesions trigger a range of cellular responses including DNA repair, cell cycle arrest, altered gene expression and cell death, which help to limit the mutagenic effects of such DNA damage. RNA processing factors are increasingly being recognised as important targets of DNA damage signalling, with roles in the regulation of gene expression and also more directly in the promotion of DNA repair. In this study, we have used a Xenopus laevis egg extract system to analyse the DNA damage-dependent phosphorylation of a putative RNA export factor, Cip29. We have found that Cip29 is rapidly phosphorylated in response to DNA double-strand breaks in this experimental system. We show that the DNA damage-inducible modification of Cip29 is dependent on the activity of the key double-strand break response kinase, ATM, and we have identified a conserved serine residue as a damage-dependent phosphorylation site. Finally, we have determined that Cip29 is not required for efficient DNA end-joining in egg extracts. Taken together, these data identify Cip29 as a novel target of the DNA damage response and suggest that the damage-dependent modification of Cip29 may relate to a role in the regulation of gene expression after DNA damage.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antibodies / immunology
  • Ataxia Telangiectasia Mutated Proteins / metabolism
  • Checkpoint Kinase 1 / metabolism
  • DNA Breaks, Double-Stranded
  • DNA Repair*
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism*
  • Mutagenesis, Site-Directed
  • Ovum / metabolism
  • Phosphorylation
  • Plasmids / metabolism
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / immunology
  • Recombinant Proteins / isolation & purification
  • Xenopus / growth & development
  • Xenopus / metabolism*
  • Xenopus Proteins / genetics
  • Xenopus Proteins / metabolism*

Substances

  • Antibodies
  • DNA-Binding Proteins
  • Recombinant Proteins
  • Xenopus Proteins
  • Atr protein, Xenopus
  • Ataxia Telangiectasia Mutated Proteins
  • Checkpoint Kinase 1

Grants and funding

JH was supported by DTP studentship funding from the Biotechnology and Biological Sciences Research Council (BBSRC grant BB\F0171111) (http://www.bbsrc.ac.uk/). Work was supported by North West Cancer Research (grant CR869 to HDL) (http://www.nwcr.org/) and Dowager Countess Eleanor Peel Trust Minor Medical Grant (http://www.peeltrust.com/) (HDL). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.