Faculty of Pharmaceutical Sciences, Tokushima Bunri University, Tokushima, 770-8514, Japan.
Department of Pharmaceutical Microbiology, Faculty of Life Sciences, Kumamoto University, Kumamoto, 862-0973, Japan.
Biochem Biophys Res Commun. 2021 Jun 11;557:273-279. doi: 10.1016/j.bbrc.2021.03.173. Epub 2021 Apr 8.
Recently, the novel coronavirus (SARS-CoV-2), which has spread from China to the world, was declared a global public health emergency, which causes lethal respiratory infections. Acetylation of several proteins plays essential roles in various biological processes, such as viral infections. We reported that the nucleoproteins of influenza virus and Zaire Ebolavirus were acetylated, suggesting that these modifications contributed to the molecular events involved in viral replication. Similar to influenza virus and Ebolavirus, the coronavirus also contains single-stranded RNA, as its viral genome interacts with the nucleocapsid (N) proteins. In this study, we report that SARS-CoV and SARS-CoV-2 N proteins are strongly acetylated by human histone acetyltransferases, P300/CBP-associated factor (PCAF), and general control nonderepressible 5 (GCN5), but not by CREB-binding protein (CBP) in vitro. Liquid chromatography-mass spectrometry analyses identified 2 and 12 acetyl-lysine residues from SARS-CoV and SARS-CoV-2 N proteins, respectively. Particularly in the SARS-CoV-2 N proteins, the acetyl-lysine residues were localized in or close to several functional sites, such as the RNA interaction domains and the M-protein interacting site. These results suggest that acetylation of SARS-CoV-2 N proteins plays crucial roles in their functions.
最近，从中国传播到世界各地的新型冠状病毒（SARS-CoV-2）被宣布为全球公共卫生紧急事件，可引起致命的呼吸道感染。几种蛋白质的乙酰化在各种生物过程中起着重要作用，如病毒感染。我们曾报道流感病毒和扎伊尔埃博拉病毒的核蛋白被乙酰化，表明这些修饰有助于病毒复制所涉及的分子事件。与流感病毒和埃博拉病毒类似，冠状病毒也含有单链 RNA，因为其病毒基因组与核衣壳（N）蛋白相互作用。在这项研究中，我们报告 SARS-CoV 和 SARS-CoV-2 N 蛋白在体外被人类组蛋白乙酰转移酶 P300/CBP 相关因子（PCAF）和一般控制非阻遏 5（GCN5）强烈乙酰化，但不能被 CREB 结合蛋白（CBP）乙酰化。液相色谱-质谱分析分别从 SARS-CoV 和 SARS-CoV-2 N 蛋白中鉴定出 2 个和 12 个乙酰化赖氨酸残基。特别是在 SARS-CoV-2 N 蛋白中，乙酰化赖氨酸残基定位于或靠近几个功能位点，如 RNA 相互作用域和 M 蛋白相互作用位点。这些结果表明，SARS-CoV-2 N 蛋白的乙酰化在其功能中起着关键作用。
