Mass spectrometry-based quantification of the cellular response to ultraviolet radiation in HeLa cells

PLoS One. 2017 Nov 20;12(11):e0186806. doi: 10.1371/journal.pone.0186806. eCollection 2017.

Abstract

Ultraviolet (UV) irradiation is a common form of DNA damage that can cause pyrimidine dimers between DNA, which can cause gene mutations, even double-strand breaks and threaten genome stability. If DNA repair systems default their roles at this stage, the organism can be damaged and result in disease, especially cancer. To better understand the cellular response to this form of damage, we applied highly sensitive mass spectrometry to perform comparative proteomics of phosphorylation in HeLa cells. A total of 4367 phosphorylation sites in 2100 proteins were identified, many of which had not been reported previously. Comprehensive bioinformatics analysis revealed that these proteins were involved in many important biological processes, including signaling, localization and cell cycle regulation. The nuclear pore complex, which is very important for RNA transport, was changed significantly at phosphorylation level, indicating its important role in response to UV-induced cellular stress. Protein-protein interaction network analysis and DNA repair pathways crosstalk were also examined in this study. Proteins involved in base excision repair, nucleotide repair and mismatch repair changed their phosphorylation pattern in response to UV treatment, indicating the complexity of cellular events and the coordination of these pathways. These systematic analyses provided new clues of protein phosphorylation in response to specific DNA damage, which is very important for further investigation. And give macroscopic view on an overall phosphorylation situation under UV radiation.

MeSH terms

  • DNA Damage / radiation effects*
  • DNA Repair / radiation effects*
  • HeLa Cells / radiation effects
  • Humans
  • Mass Spectrometry
  • Phosphorylation / radiation effects
  • Protein Interaction Maps / radiation effects
  • Pyrimidine Dimers / radiation effects*
  • Ultraviolet Rays

Substances

  • Pyrimidine Dimers

Grants and funding

The work was supported by the following: National Basic Research Program of China (2015CB910600 to YH); National Natural Science Foundation of China (31210103904 to YH), Natural Science Foundation of Zhejiang Province (LY16C050003 to HX), National Key R&D Program of China (2017YFA0503900 to YH), and The Science Technology Department, Zhejiang Province (KN20160607 to HX). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.