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The membrane-associated ubiquitin ligases MARCH2 and MARCH3 target TIM-1 to limit Zika virus infection

Cellular & Molecular Immunology volume 22, pages 1032–1044 (2025)Cite this article

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A Comment to this article was published on 28 November 2025

Abstract

T-cell immunoglobulin mucin family member-1 (TIM-1, also known as HAVCR1/KIM-1) is a transmembrane glycoprotein that has been reported to act as an entry receptor for multiple flaviviruses including Zika virus (ZIKV). The post-translational regulation of TIM-1 and its effects on ZIKV infection are unclear. In this study, we identified the membrane-associated RING-CH-type finger (MARCH) E3 ubiquitin ligase family members MARCH2 and MARCH3 as critical negative regulators of TIM-1 under physiological conditions. MARCH2 and MARCH3 associate with TIM-1 and mediate its K48-linked polyubiquitination at K338 and K346 respectively, leading to subsequent proteasomal degradation. While deficiency of either MARCH2 or MARCH3 modestly increases TIM-1 levels and enhances ZIKV infectivity, double knockout of MARCH2/3 has a more dramatic effect. Double knockout of MARCH2/3 increased ZIKV infectivity in wild-type but not TIM-1 knockout cells, and reconstitution of TIM-1K338R/K346R into TIM-1-deficient cells increases ZIKV infectivity to a higher degree than reconstitution with wild-type TIM-1. Knockout of either MARCH2 or MARCH3 increased ZIKV infectivity and pathogenesis in mice, whereas double knockout of MARCH2/3 has a more dramatic effect. These findings suggest that MARCH2 and MARCH3 target TIM-1 for K48-linked polyubiquitination and proteasomal degradation, thereby acting as redundant host restriction factors to limit ZIKV infection and pathogenesis.

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Acknowledgements

We thank Wen-Hua Xu, Xu Chen, and other members of our laboratory for technical help and discussions. This work was supported by grants from the State Key R&D Program of China (2024YFA1306500, 2022YFA1304900), the National Natural Science Foundation of China (32188101), the Major Project of Guangzhou National Laboratory (GZNL2024A01014, GZNL2024A01016), the Fundamental Research Funds for the Central Universities (2042022dx0003), and Natural Science Foundation of Wuhan (2024040701010031).

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Authors and Affiliations

  1. Department of Infectious Diseases, Zhongnan Hospital of Wuhan University; Hubei Provincial Research Center for Basic Biological Sciences; Medical Research Institute; Frontier Science Center for Immunology and Metabolism; Taikang Center for Life and Medical Sciences; Wuhan University, Wuhan, 430071, China

    Qi Zhang, Zhen-Wu Ma, Hui-Fang Li, Jia-Qing Zeng, Hong-Bing Shu & Shu Li

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Contributions

QZ, SL, and H-BS designed research; QZ, Z-WM, H-FL, and J-QZ performed research; QZ, SL, and H-BS analyzed data; and QZ, SL, and H-BS wrote the paper.

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Correspondence to Hong-Bing Shu or Shu Li.

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The authors declare no competing interests. Dr. H.-B.S. is editorial board member of Cellular & Molecular Immunology, but he has not been involved in the peer review or the decision-making of the article.

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Zhang, Q., Ma, ZW., Li, HF. et al. The membrane-associated ubiquitin ligases MARCH2 and MARCH3 target TIM-1 to limit Zika virus infection. Cell Mol Immunol 22, 1032–1044 (2025). https://doi.org/10.1038/s41423-025-01334-2

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