Mass spectrometry-based quantification of the cellular response to ultraviolet radiation in HeLa cells.
Mass spectrometry-based quantification of the cellular response to ultraviolet radiation in HeLa cells.
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DOI：
10.1371/journal.pone.0186806
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发表时间：
2017
期刊：
影响因子：
3.7
通讯作者：
Hua Y
中科院分区：
文献类型：
--
作者：
Xu H;Chen X;Ying N;Wang M;Xu X;Shi R;Hua Y
紫外线（UV）照射是一种常见的DNA损伤形式，可导致DNA之间的嘧啶二聚体，从而导致基因突变，甚至双链断裂，威胁基因组稳定性。如果DNA修复系统在这个阶段没有发挥作用，生物体可能会受到损害并导致疾病，特别是癌症。为了更好地了解细胞对这种形式的损伤的反应，我们应用高灵敏度质谱法对HeLa细胞中的磷酸化进行比较蛋白质组学研究。在2100个蛋白质中共鉴定出4367个磷酸化位点，其中许多是以前没有报道过的。综合生物信息学分析表明，这些蛋白质参与了许多重要的生物学过程，包括信号传导、定位和细胞周期调控。核孔复合体是RNA转运的重要组成部分，在磷酸化水平上发生了显著变化，表明其在细胞响应紫外胁迫中的重要作用。蛋白质-蛋白质相互作用网络分析和DNA修复途径的串扰也在这项研究中进行了检查。参与碱基切除修复、核苷酸修复和错配修复的蛋白质改变了它们的磷酸化模式以响应UV处理，表明细胞事件的复杂性和这些途径的协调性。这些系统分析为特异性DNA损伤后蛋白质磷酸化反应提供了新的线索，对进一步研究具有重要意义。并给出了紫外光照射下整体磷酸化情况的宏观视图。
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.
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