Gu Lin, Wang Zheng-Jie, Zhang Xin-Ran, Liu Yu, Zhao Ming, Jiang Shao-Zhen, Pan Jie, Yuan Ying, Cai Hong, Zhou Tao, Li Tao, Li Ting-Ting, Xue Wen
Nanhu Laboratory, National Center of Biomedical Analysis, Beijing, 100850, China.
Nanhu Laboratory, National Center of Biomedical Analysis, Beijing, 100850, China; School of Basic Medical Sciences, Fudan University, Shanghai, 200032, China.
Biochem Biophys Res Commun. 2025 Apr 5;756:151594. doi: 10.1016/j.bbrc.2025.151594. Epub 2025 Mar 6.
The COVID-19 pandemic, caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has led to a global public health crisis. The nucleocapsid (N) protein plays a pivotal role in a variety of biological processes in the life cycle of SARS-CoV-2, such as viral assembly. In this study, we investigated the liquid-liquid phase separation (LLPS) capacity of the N protein of seven SARS-CoV-2 strains, including the variants of concern (VOC) and interest (VOI), and its impact on viral replication. Using bioinformatic tools, we analyzed 11,433,558 complete genomes of SARS-CoV-2 and revealed a high degree of sequence conservation of N gene. While all the seven N proteins could undergo LLPS with RNA, the mutations in N impair its capacity of LLPS. With a SARS-CoV-2 trans-complementation system, we showed that SARS-CoV-2 variants carrying mutated N proteins exhibit impaired replication, highlighting the importance of LLPS of N in viral replication. We further demonstrated that (-)-gallocatechin gallate (GCG) efficiently inhibits the LLPS of N proteins and significantly suppresses the replication of different SARS-CoV-2 strains. Thus, our findings indicate that targeting the N-LLPS could be a viable strategy for the development of antiviral treatments against various SARS-CoV-2 strains, including those yet to emerge.
由严重急性呼吸综合征冠状病毒2（SARS-CoV-2）引起的COVID-19大流行已导致全球公共卫生危机。核衣壳（N）蛋白在SARS-CoV-2生命周期的各种生物学过程中起着关键作用，例如病毒组装。在本研究中，我们调查了七种SARS-CoV-2毒株（包括关注变异株（VOC）和感兴趣变异株（VOI））的N蛋白的液-液相分离（LLPS）能力及其对病毒复制的影响。使用生物信息学工具，我们分析了11433558个SARS-CoV-2的完整基因组，并揭示了N基因的高度序列保守性。虽然所有七种N蛋白都可以与RNA发生液-液相分离，但N中的突变会损害其液-液相分离能力。通过SARS-CoV-2转互补系统，我们表明携带突变N蛋白的SARS-CoV-2变异株表现出复制受损，突出了N的液-液相分离在病毒复制中的重要性。我们进一步证明，（-）-没食子儿茶素没食子酸酯（GCG）有效抑制N蛋白的液-液相分离，并显著抑制不同SARS-CoV-2毒株的复制。因此，我们的研究结果表明，靶向N的液-液相分离可能是开发针对各种SARS-CoV-2毒株（包括尚未出现的毒株）的抗病毒治疗方法的可行策略。
