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

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

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
Available online: 08 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

Electronic Supplementary Material

10.1016j.virs.2026.07.002-ESM.docx
References
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The C-terminal T234 residue of VP3 is required for KREMEN1 receptor-dependent infectivity and pathogenesis of coxsackieviruses A10 and A8

Corresponding author: Chao Zhang, chao_zhang@fudan.edu.cn

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

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.

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

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