Arce Rodrigo, Soñora Martín, Andreu-Moreno Iván, Moreno Pilar, Moratorio Gonzalo, Sanjuán Rafael
Virus Evolution Laboratory, Institute for Integrative Systems Biology (I2SysBio), Universitat de València-CSIC, Paterna, Spain.
Molecular Virology Laboratory, Center for Nuclear Research, Faculty of Sciences, Universidad de la República, Montevideo, Uruguay.
Sci Rep. 2025 Mar 10;15(1):8314. doi: 10.1038/s41598-025-92289-x.
Coxsackievirus B3 (CVB3), a member of the Enterovirus genus within the Picornaviridae family, has emerged as a key model for studying viral evolution and pathogenesis. Although traditionally considered obligate lytic viruses, recent research reveals that enteroviruses can also be released non-lytically within extracellular vesicles (EVs). This study explores the impact of mutations at position 63 of the VP3 capsid protein on CVB3 fitness and release mechanisms by substituting asparagine at this position with aromatic, charged, and aliphatic amino acids. We show that mutations at position 63 significantly affect viral release mechanisms and viral spread in cell culture. Specifically, aromatic mutations (N63H, N63Y, N63F, N63W) and the N63D mutation reduce the release of membrane-associated viral particles, while aromatic residues increase viral spread in cell culture and plaque size under specific conditions. These findings suggest that N63 mutations alter protomer interactions, influencing viral release, spread, and plaque formation, providing insights into the molecular mechanisms of CVB3 egress.
柯萨奇病毒B3（CVB3）是小核糖核酸病毒科肠道病毒属的成员，已成为研究病毒进化和发病机制的关键模型。尽管传统上认为肠道病毒是专性裂解病毒，但最近的研究表明，肠道病毒也可以在细胞外囊泡（EVs）内以非裂解方式释放。本研究通过用芳香族、带电荷和脂肪族氨基酸取代VP3衣壳蛋白第63位的天冬酰胺，探讨该位点突变对CVB3适应性和释放机制的影响。我们发现，第63位的突变显著影响病毒的释放机制和在细胞培养中的传播。具体而言，芳香族突变（N63H、N63Y、N63F、N63W）和N63D突变减少了膜相关病毒颗粒的释放，而芳香族残基在特定条件下增加了病毒在细胞培养中的传播和蚀斑大小。这些发现表明，N63突变改变了原体相互作用，影响病毒释放、传播和蚀斑形成，为CVB3释放的分子机制提供了见解。
