The C-terminal T234 residue of VP3 is required for KREMEN1 receptor-dependent infectivity and pathogenesis of coxsackieviruses A10 and A8

Xingyu Yan; Zeyu Liu; Kexin Liu; Zhenlin Yang; Jianxing Wang; Chao Zhang

doi:10.1016/j.virs.2026.07.002

August 2026

. doi: 10.1016/j.virs.2026.07.002

Citation: Xingyu Yan, Zeyu Liu, Kexin Liu, Zhenlin Yang, Jianxing Wang, Chao Zhang. The C-terminal T234 residue of VP3 is required for KREMEN1 receptor-dependent infectivity and pathogenesis of coxsackieviruses A10 and A8 .VIROLOGICA SINICA, 2026, 41(4) : 882-897. http://dx.doi.org/10.1016/j.virs.2026.07.002

VP3蛋白C端T234残基是柯萨奇病毒A10和A8通过KREMEN1受体依赖方式实现感染性及致病性所必需的关键位点

阎星雨 ^a ,
刘则宇 ^a ,
刘可歆 ^a ,
杨振霖 ^b,c ,
王建醒 ^d ,
张超 ^a,,

a 复旦大学上海市重大传染病和生物安全研究院;
b 复旦大学附属中山医院呼吸科, 上海市肺部炎症与损伤重点实验室;
c 上海市合成免疫工程技术研究中心;
d 山东省疾病预防控制中心

通讯作者： 张超, chao_zhang@fudan.edu.cn
收稿日期： 2026-05-20
录用日期： 2026-07-06

摘要

柯萨奇病毒A10（CVA10）是手足口病的主要病原体之一，利用KREMEN1（KRM1）作为其细胞受体。虽然前期的研究确定了VP2残基K140是KRM1依赖性肠道病毒与KRM1结合的通用锚点，但其他受体界面残基的功能意义尚不清楚。本研究通过结构导向的定点突变，发现VP3蛋白C端完全保守的T234残基是CVA10感染所必需的。T234A突变不影响病毒颗粒组装，但完全消除了与KRM1结合和细胞吸附。有趣的是，这种需求表现出显著的病毒特异性：同源残基对CVA8至关重要，但对其他利用KRM1的肠道病毒（包括CVA2–CVA6和CVA12）并非必需。T234A突变显著减弱了CVA10和CVA8在新生小鼠中的致病性。此外，CVA8-T234A突变株作为减毒疫苗，能够完全保护新生小鼠免受致死剂量CVA8的攻毒。我们的发现建立了一个模型：KRM1结合依赖保守的VP2-K140锚点以及病毒特异性次级残基，其中VP3-T234是CVA10和CVA8的关键决定位点。这些见解加深了我们对肠道病毒-受体相互作用的理解，并为疫苗开发提供了新方向。
- 手足口病
- , 肠道病毒
- , KREMEN1受体
- , 疫苗

The C-terminal T234 residue of VP3 is required for KREMEN1 receptor-dependent infectivity and pathogenesis of coxsackieviruses A10 and A8

Xingyu Yan ^a ,
Zeyu Liu ^a ,
Kexin Liu ^a ,
Zhenlin Yang ^b,c ,
Jianxing Wang ^d ,
Chao Zhang ^a,,

a. Shanghai Institute of Infectious Disease and Biosecurity, Fudan University, Shanghai 200032, China;
b. Shanghai Key Laboratory of Lung Inflammation and Injury, Department of Pulmonary Medicine, Zhongshan Hospital, Fudan University, Shanghai 200032, China;
c. Shanghai Engineering Research Center for Synthetic Immunology, Shanghai 200032, China;
d. Shandong Center for Disease Control and Prevention, Jinan 250014, China

Corresponding author: Chao Zhang, chao_zhang@fudan.edu.cn
Received Date: 20 May 2026
Accepted Date: 06 July 2026

Abstract

Coxsackievirus A10 (CVA10) is a major causative agent of hand, foot and mouth disease and utilizes KREMEN1 (KRM1) as its cellular receptor. While our previous work identifies VP2 residue K140 as a universal anchor for KRM1 binding among KRM1-utilizing enteroviruses, the functional significance of other receptor-interface residues remains poorly characterized. Here, through structure-guided mutagenesis, we demonstrate that VP3-T234, a completely conserved residue at the C-terminus of VP3, is essential for CVA10 infectivity. The T234A mutation does not affect virion assembly but abolishes both KRM1 binding and cellular attachment. Interestingly, this requirement shows remarkable virus specificity: the homologous residue is critical for CVA8, but is not required for other KRM1-utilizing enteroviruses including CVA2-CVA6 and CVA12. The T234A mutation significantly attenuates the pathogenesis of both CVA10 and CVA8 in neonatal mice. Moreover, the CVA8-T234A mutant provides complete protection as an attenuated vaccine against lethal CVA8 challenge. Our findings establish a model wherein KRM1 engagement relies on the conserved VP2-K140 anchor complemented by virus-specific secondary residues, with VP3-T234 representing a key determinant for CVA10 and CVA8. These insights advance our understanding of enterovirus-receptor interactions and provide new directions for vaccine development.
- Hand, Foot, and mouth disease (HFMD)
- , Enterovirus
- , KREMEN1 receptor
- , Vaccine

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Electronic Supplementary Material

10.1016j.virs.2026.07.002-ESM.docx