Wang Chunhua, Yu Tingyu, Xie Moyue, Zhan Dongang, Zhang Shuaijie, Chai Wenqi, Zhao Jianzhong, Yin Lijuan, Yang Yang, Shen Liang, Xia Ying
Department of Central Laboratory, Xiangyang Central Hospital, Affiliated Hospital of Hubei University of Arts and Science, Xiangyang, 441021, Hubei Province, China.
Department of Clinical Laboratory, Xiangyang No. 1 People's Hospital, Hubei University of Medicine, Xiangyang, 441000, Hubei Province, China.
BMC Microbiol. 2025 Aug 1;25(1):474. doi: 10.1186/s12866-025-04226-7.
The nucleocapsid protein (N protein) of SARS-CoV-2 is highly conserved in viral evolution and serves as the primary structural protein in viral infection, being the most abundant in viral particles. The N protein is highly immunogenic and plays a key role in the processes of viral infection and replication, making it of significant research value in both basic studies and clinical applications.
To further investigate the functions of SARS-CoV-2 N protein, the Matchmaker Gold Yeast Two-Hybrid System was used to identify potential interacting partners of the N protein in human peripheral blood mononuclear cells (PBMCs). Through this approach, we identified 11 host proteins that might interact with the SARS-CoV-2 N protein. We further validated the interaction between the N protein and two host proteins, RNF2 and ARL15, which showed the highest positive clone rates at the cellular level. We also predicted the critical amino acid residues mediating the interaction of the N protein with RNF2 or ARL15. Additionally, we explored the impact of these two host proteins on coronavirus replication. Functional analysis of all the 11 host proteins revealed their involvement in ribosome biogenesis, antigen processing and presentation, as well as various signaling pathways such as JAK-STAT.
This study further enriches the understanding of interactions between the SARS-CoV-2 N protein and host, providing important theoretical insights for the deeper understanding of viral pathogenesis and the development of antiviral strategies.
严重急性呼吸综合征冠状病毒2（SARS-CoV-2）的核衣壳蛋白（N蛋白）在病毒进化过程中高度保守，是病毒感染中的主要结构蛋白，在病毒颗粒中含量最为丰富。N蛋白具有高度免疫原性，在病毒感染和复制过程中起关键作用，这使其在基础研究和临床应用中都具有重要的研究价值。
为进一步研究SARS-CoV-2 N蛋白的功能，采用Matchmaker Gold酵母双杂交系统在人外周血单核细胞（PBMCs）中鉴定N蛋白的潜在相互作用伙伴。通过该方法，我们鉴定出11种可能与SARS-CoV-2 N蛋白相互作用的宿主蛋白。我们进一步验证了N蛋白与两种宿主蛋白RNF2和ARL15之间的相互作用，这两种蛋白在细胞水平上显示出最高的阳性克隆率。我们还预测了介导N蛋白与RNF2或ARL15相互作用的关键氨基酸残基。此外，我们探讨了这两种宿主蛋白对冠状病毒复制的影响。对所有11种宿主蛋白的功能分析表明，它们参与核糖体生物合成、抗原加工和呈递，以及各种信号通路，如JAK-STAT。
本研究进一步丰富了对SARS-CoV-2 N蛋白与宿主之间相互作用的理解，为更深入了解病毒发病机制和开发抗病毒策略提供了重要的理论见解。
