Doxorubicin-induced DNA Damage Causes Extensive Ubiquitination of Ribosomal Proteins Associated with a Decrease in Protein Translation

Mol Cell Proteomics. 2018 Dec;17(12):2297-2308. doi: 10.1074/mcp.RA118.000652. Epub 2018 Feb 8.

Abstract

Protein posttranslational modifications (PTMs) play a central role in the DNA damage response. In particular, protein phosphorylation and ubiquitination have been shown to be essential in the signaling cascade that coordinates break repair with cell cycle progression. Here, we performed whole-cell quantitative proteomics to identify global changes in protein ubiquitination that are induced by DNA double-strand breaks. In total, we quantified more than 9,400 ubiquitin sites and found that the relative abundance of ∼10% of these sites was altered in response to DNA double-strand breaks. Interestingly, a large proportion of ribosomal proteins, including those from the 40S as well as the 60S subunit, were ubiquitinated in response to DNA damage. In parallel, we discovered that DNA damage leads to the inhibition of ribosome function. Taken together, these data uncover the ribosome as a major target of the DNA damage response.

Keywords: Cell cycle*; Mass Spectrometry; Pathway Analysis; Ribosomes*; Ubiquitin.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line, Tumor
  • Cysteine Proteinase Inhibitors / metabolism
  • DNA Breaks, Double-Stranded*
  • Doxorubicin / pharmacology*
  • G2 Phase / physiology
  • Humans
  • Leupeptins / metabolism
  • Mass Spectrometry
  • Nuclear Proteins / metabolism
  • Nucleolin
  • Nucleophosmin
  • Phosphoproteins / metabolism
  • Phosphorylation
  • Protein Biosynthesis / physiology*
  • Protein Processing, Post-Translational*
  • RNA-Binding Proteins / metabolism
  • Ribosomal Proteins / metabolism*
  • Ribosomes / metabolism
  • Signal Transduction
  • Ubiquitin / metabolism
  • Ubiquitinated Proteins / metabolism
  • Ubiquitination / physiology*

Substances

  • Cysteine Proteinase Inhibitors
  • Leupeptins
  • Nuclear Proteins
  • Phosphoproteins
  • RNA-Binding Proteins
  • Ribosomal Proteins
  • Ubiquitin
  • Ubiquitinated Proteins
  • Nucleophosmin
  • Doxorubicin
  • benzyloxycarbonylleucyl-leucyl-leucine aldehyde