Unraveling the assembly mechanism of SADS-CoV virus nucleocapsid protein: insights from RNA binding, dimerization, and epitope diversity profiling

被引:2
作者
Zhang, Ying [1 ,2 ,3 ]
Wu, Fang [4 ]
Han, Yongyue [1 ,2 ,3 ]
Wu, Yuzhe [1 ,2 ,3 ]
Huang, Liqiu [1 ,2 ,3 ]
Huang, Yuanwei [1 ,2 ,3 ]
Yan, Di [1 ,2 ,3 ]
Jiang, Xiwen [5 ]
Ma, Jingyun [6 ]
Xu, Wei [1 ,2 ,3 ,7 ]
机构
[1] Southern Med Univ, Guangdong Prov Key Lab New Drug Screening, Guangzhou, Peoples R China
[2] Southern Med Univ, NMPA Key Lab Res & Evaluat Drug Metab, Guangzhou, Peoples R China
[3] Southern Med Univ, Sch Pharmaceut Sci, Guangdong Hong Kong Macao Joint Lab New Drug Scree, Guangzhou, Peoples R China
[4] Southern Med Univ, Affiliated Foshan Matern & Child Healthcare Hosp, Foshan, Peoples R China
[5] Guangdong Pharmaceut Univ, Sch Life Sci & Biopharmaceut, Guangzhou, Peoples R China
[6] South China Agr Univ, Coll Anim Sci, Guangzhou, Peoples R China
[7] Southern Med Univ, Key Lab Infect Dis Res South China, Minist Educ, Guangzhou, Peoples R China
基金
中国国家自然科学基金;
关键词
SADS-CoV virus; nucleocapsid protein; RNA binding; dimerization; INTRINSICALLY DISORDERED PROTEINS; CORONAVIRUS; DOMAIN; SWINE;
D O I
10.1128/jvi.00926-24
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
The swine acute diarrhea syndrome coronavirus (SADS-CoV) has caused significant disruptions in porcine breeding and raised concerns about potential human infection. The nucleocapsid (N) protein of SADS-CoV plays a vital role in viral assembly and replication, but its structure and functions remain poorly understood. This study utilized biochemistry, X-ray crystallography, and immunization techniques to investigate the N protein's structure and function in SADS-CoV. Our findings revealed distinct domains within the N protein, including an RNA-binding domain, two disordered domains, and a dimerization domain. Through biochemical assays, we confirmed that the N-terminal domain functions as an RNA-binding domain, and the C-terminal domain is involved in dimerization, with the crystal structure analysis providing visual evidence of dimer formation. Immunization experiments demonstrated that the disordered domain 2 elicited a significant antibody response. These identified domains and their interactions are crucial for viral assembly. This comprehensive understanding of the N protein in SADS-CoV enhances our knowledge of its assembly and replication mechanisms, enabling the development of targeted interventions and therapeutic strategies.IMPORTANCESADS-CoV is a porcine coronavirus that originated from a bat HKU2-related coronavirus. It causes devastating swine diseases and poses a high risk of spillover to humans. The coronavirus N protein, as the most abundant viral protein in infected cells, likely plays a key role in viral assembly and replication. However, the structure and function of this protein remain unclear. Therefore, this study employed a combination of biochemistry and X-ray crystallography to uncover distinct structural domains in the N protein, including RNA-binding domains, two disordered domains, and dimerization domains. Additionally, we made the novel discovery that the disordered domain elicited a significant antibody response. These findings provide new insights into the structure and functions of the SADS-CoV N protein, which have important implications for future studies on SADS-CoV diagnosis, as well as the development of vaccines and anti-viral drugs. SADS-CoV is a porcine coronavirus that originated from a bat HKU2-related coronavirus. It causes devastating swine diseases and poses a high risk of spillover to humans. The coronavirus N protein, as the most abundant viral protein in infected cells, likely plays a key role in viral assembly and replication. However, the structure and function of this protein remain unclear. Therefore, this study employed a combination of biochemistry and X-ray crystallography to uncover distinct structural domains in the N protein, including RNA-binding domains, two disordered domains, and dimerization domains. Additionally, we made the novel discovery that the disordered domain elicited a significant antibody response. These findings provide new insights into the structure and functions of the SADS-CoV N protein, which have important implications for future studies on SADS-CoV diagnosis, as well as the development of vaccines and anti-viral drugs.
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页数:22
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