Huang Qihong, Lin Zijia, Wu Pengju, Ni Jinfeng, Shen Yulong
CRISPR and Archaea Biology Research Center, State Key Laboratory of Microbial Technology, Microbial Technology Institute, Shandong University, Qingdao, China.
Front Microbiol. 2020 Dec 3;11:586025. doi: 10.3389/fmicb.2020.586025. eCollection 2020.
DNA damage response (DDR) in eukaryotes is largely regulated by protein phosphorylation. In archaea, many proteins are phosphorylated, however, it is unclear how the cells respond to DNA damage through global protein phosphorylation. We previously found that Δ, a Rio1 kinase homolog deletion strain of REY15A, was sensitive to UV irradiation. In this study, we showed that Δ grew faster than the wild type. Quantitative phosphoproteomic analysis of the wild type and Δ, untreated and irradiated with UV irradiation, revealed 562 phosphorylated sites (with a Ser/Thr/Tyr ratio of 65.3%/23.8%/10.9%) of 333 proteins in total. The phosphorylation levels of 35 sites of 30 proteins changed with >1.3-fold in the wild type strain upon UV irradiation. Interestingly, more than half of the UV-induced changes in the wild type did not occur in the Δ strain, which were mainly associated with proteins synthesis and turnover. In addition, a protein kinase and several transcriptional regulators were differentially phosphorylated after UV treatment, and some of the changes were dependent on Rio1. Finally, many proteins involved in various cellular metabolisms exhibited Riol-related and UV-independent phosphorylation changes. Our results suggest that Rio1 is involved in the regulation of protein recycling and signal transduction in response to UV irradiation, and plays regulatory roles in multiple cellular processes in .
真核生物中的DNA损伤反应（DDR）在很大程度上受蛋白质磷酸化调控。在古细菌中，许多蛋白质会发生磷酸化，然而，目前尚不清楚细胞如何通过全局蛋白质磷酸化对DNA损伤作出反应。我们之前发现，REY15A的Rio1激酶同源缺失菌株Δ对紫外线照射敏感。在本研究中，我们发现Δ的生长速度比野生型更快。对未经处理和经紫外线照射的野生型和Δ进行定量磷酸化蛋白质组分析，总共揭示了333种蛋白质的562个磷酸化位点（丝氨酸/苏氨酸/酪氨酸比例为65.3%/23.8%/10.9%）。在野生型菌株中，紫外线照射后30种蛋白质的35个位点的磷酸化水平变化超过1.3倍。有趣的是，野生型中超过一半的紫外线诱导变化在Δ菌株中未发生，这些变化主要与蛋白质合成和周转相关。此外，紫外线处理后一种蛋白激酶和几种转录调节因子发生了差异磷酸化，其中一些变化依赖于Rio1。最后，许多参与各种细胞代谢的蛋白质表现出与Rio1相关且与紫外线无关的磷酸化变化。我们的结果表明，Rio1参与了紫外线照射响应中蛋白质循环和信号转导的调控，并在REY15A的多个细胞过程中发挥调节作用。
