African swine fever virus pD345L negatively regulates the cGAS-STING signaling pathway by inhibiting the binding of STING and IRF3

doi:10.1016/j.jia.2026.04.005

Advanced Online Publication | Current Issue | Archive | Adv Search

Xiaohong Liu^1*, Xiaoping He^1*, Hongyang Liu¹, Yutong Wang¹, Siqi Dong¹, Hanyu Wu¹, Yi Zeng¹, Tingting Li^{1, 2}, Zhaoxia Zhang^{1, 2}, Jiangnan Li^{1, 2}, Changjiang Weng^{1, 2#}, Li Huang^{1, 2#}

¹ National African Swine Fever Para-reference Laboratory, State Key Laboratory of Animal Disease Control and Prevention, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150069, China

²Heilongjiang Provincial Key Laboratory of Veterinary Immunology, Harbin 150069, China

Highlights

Ÿ ASFV pD345L inhibits IFN‑I production by negatively regulating cGAS-STING pathway.

Ÿ ASFV pD345L inhibits the formation of the STING-IRF3 signaling complex.

Ÿ The C‑terminal domain of ASFV pD345L is responsible for its inhibitory effect on IFN‑I production.

Abstract
References

Download: PDF in ScienceDirect
Export: BibTeX | EndNote (RIS)

摘要

非洲猪瘟（African swine fever, ASF）是由非洲猪瘟病毒（African swine fever virus, ASFV）引起的家猪高度传染性出血性疾病，致死率接近 100%，对全球养猪业造成毁灭性打击。ASFV 作为大型双链 DNA 病毒，感染宿主细胞后其基因组可被胞质 DNA 识别受体环 GMP-AMP 合酶（cyclic GMP-AMP synthase, cGAS）感知，进而激活 cGAS-STING（stimulator of interferon genes）信号通路，诱导 I 型干扰素（type I interferon, IFN-I）等细胞因子产生，启动宿主天然免疫应答。为实现高效感染与持续传播，ASFV 已进化出多种策略靶向该通路进行免疫逃逸，但其具体分子机制尚未完全阐明。本研究旨在探究 ASFV 编码蛋白 pD345L 对 cGAS-STING 通路的调控作用及分子机制，为揭示 ASFV 的免疫逃逸策略提供实验依据。本研究通过采用双荧光素酶报告基因检测系统，检测 pD345L 对 cGAS-STING 通路下游启动子活性的影响，结果显示，ASFV pD345L 以剂量依赖性方式显著抑制 cGAS-STING 通路介导的 IFN-β、IFN-α 及 ISG54 启动子活性；通过实时荧光定量聚合酶链式反应（qRT-PCR）检测 pD345L 对 cGAS-STING 诱导的 IFN-I 相关基因 mRNA 表达水平的调控作用，结果证实，pD345L 可明显降低 cGAS-STING 诱导的 IFNβ、ISG54、ISG56 基因 mRNA 表达水平。利用免疫共沉淀技术，分析 pD345L 与 cGAS-STING 通路关键分子的相互作用，结果表明，pD345L 可分别与 STING 和 IRF3 发生相互作用，其中pD345L的C末端是其发挥作用的关键结构域；进一步研究发现，pD345L 的相互作用可阻断 STING 与 IRF3 信号复合物的形成，进而抑制 IRF3 的磷酸化修饰及核转位过程，最终导致 IFN-I 产生的显著下调。研究首次证实 ASFV 编码蛋白 pD345L 是 cGAS-STING 信号通路的新型负调控因子，其通过靶向结合 STING 与 IRF3，阻断二者信号复合物的组装，从而负向调控 cGAS-STING 通路介导的 IFN-I 免疫应答。该研究揭示了 ASFV 逃逸宿主天然免疫的全新分子机制，不仅丰富了 ASFV 免疫逃逸的理论体系，也为开发针对 ASFV 的新型疫苗设计、抗病毒药物靶点筛选及 ASF 防控策略制定提供了关键实验依据和理论支撑。

Abstract

African swine fever (ASF) is a highly contagious and hemorrhagic disease caused by African swine fever virus (ASFV), with a mortality rate approaching 100% in domestic pigs. ASFV is a large DNA virus, and its genome can be recognized by the cytoplasmic DNA sensor cyclic GMP-AMP synthase (cGAS) following infection to trigger the production of type I interferon (IFN-I) through the cGAS-STING signaling pathway. To establish productive infection, ASFV encodes multiple proteins to negatively regulate the cGAS-STING pathway and inhibit the expression of IFN-I. However, the molecular mechanisms by which ASFV proteins negatively regulate cGAS-STING signaling pathway remain incompletely elucidated. Through screening ASFV-encoded proteins, we found that pD345L significantly inhibits IFN-I production. Furthermore, we demonstrate that ASFV pD345L inhibits the promoter activities of Interferon-β (IFN-β)-, Interferon-α (IFN-α)-, interferon-stimulated gene (ISG)-54-Luciferase (Luc), as well as the mRNA levels of IFN-β, ISG-54, ISG-56 induced by cGAS-STING in a dose-dependent manner. Moreover, our findings reveal that ASFV pD345L interacts with both stimulator of interferon genes (STING) and interferon regulatory factor 3 (IRF3), thereby disrupting the formation of the STING-IRF3 complex. This interaction leads to impaired IRF3 phosphorylation and nuclear translocation, ultimately suppressing the production of IFN-I. Collectively, our findings reveal that ASFV pD345L functions as a negative regulator of the cGAS-STING signaling pathway to inhibit IFN-I production, thereby facilitating the viral evasion of the host innate immune response.

Keywords: African swine fever virus pD345L cGAS-STING signaling IFN-I

Online: 07 April 2026

Fund:

This study was supported by the National Natural Science Foundation of China (32322081 and 32270156), Natural Science Foundation of Heilongjiang Province of China (ZD2025C009), Innovation Program of Chinese Academy of Agricultural Sciences (CAAS-CSLPDCP-202401, CAAS-CSLPDCP-2023002), the Central Public-interest Scientific Institution Basal Research Fund (Y2025YC123).

About author: #Correspondence Changjiang Weng, E-mail: wengchangjiang@caas.cn; Li Huang, E-mail: huangli02@caas.cn *Xiaohong Liu and Xiaoping He contributed equally to this work.

	Service
	E-mail this article
	Add to citation manager
	E-mail Alert
	RSS
	Articles by authors

Cite this article:

Xiaohong Liu, Xiaoping He, Hongyang Liu, Yutong Wang, Siqi Dong, Hanyu Wu, Yi Zeng, Tingting Li, Zhaoxia Zhang, Jiangnan Li, Changjiang Weng, Li Huang. 2026. African swine fever virus pD345L negatively regulates the cGAS-STING signaling pathway by inhibiting the binding of STING and IRF3. Journal of Integrative Agriculture, Doi:10.1016/j.jia.2026.04.005

Abe T and Barber G N. 2014. Cytosolic-DNA-mediated, STING-dependent proinflammatory gene induction necessitates canonical NF-kappaB activation through TBK1. Journal of virology, 88, 5328-5341.

Chapman D A, Darby A C, Da Silva M, Upton C, Radford A D and Dixon L K. 2011. Genomic analysis of highly virulent Georgia 2007/1 isolate of African swine fever virus. Emerging Infectious Diseases, 17, 599-605.

Chathuranga K and Lee J S. 2023. African Swine Fever Virus (ASFV): Immunity and Vaccine Development. Vaccines (Basel), 11, 199.

Chen C and Xu P. 2023. Cellular functions of cGAS-STING signaling. Trends in Cell Biology, 33, 630-648.

Chen H, Wang Z, Gao X, Lv J, Hu Y, Jung Y S, Zhu S, Wu X, Qian Y and Dai J. 2022. ASFV pD345L protein negatively regulates NF-κB signalling by inhibiting IKK kinase activity. Veterinary Research, 53, 32.

Chen Q, Sun L and Chen Z J. 2016. Regulation and function of the cGAS-STING pathway of cytosolic DNA sensing. Nature Immunology, 17, 1142-1149.

Collins A C, Cai H, Li T, Franco L H, Li X D, Nair V R, Scharn C R, Stamm C E, Levine B, Chen Z J and Shiloh M U. 2015. Cyclic GMP-AMP Synthase Is an Innate Immune DNA Sensor for Mycobacterium tuberculosis. Cell Host Microbe, 17, 820-828.

de Villiers E P, Gallardo C, Arias M, da Silva M, Upton C, Martin R and Bishop R P. 2010. Phylogenomic analysis of 11 complete African swine fever virus genome sequences. Virology, 400, 128-136.

Dixon L K, Chapman D A, Netherton C L and Upton C. 2013. African swine fever virus replication and genomics. Virus Research, 173, 3-14.

Dodantenna N, Ranathunga L, Chathuranga W A G, Weerawardhana A, Cha J W, Subasinghe A, Gamage N, Haluwana D K, Kim Y, Jheong W, Poo H and Lee J S. 2022. African Swine Fever Virus EP364R and C129R Target Cyclic GMP-AMP To Inhibit the cGAS-STING Signaling Pathway. Journal of virology, 96, e0102222.

Gao Q, Yang Y, Luo Y, Chen X, Gong T, Wu D, Feng Y, Zheng X, Wang H, Zhang G, Lu G and Gong L. 2023. African Swine Fever Virus Envelope Glycoprotein CD2v Interacts with Host CSF2RA to Regulate the JAK2-STAT3 Pathway and Inhibit Apoptosis to Facilitate Virus Replication. Journal of virology, 97, e0188922.

García-Belmonte R, Pérez-Núñez D, Pittau M, Richt J A and Revilla Y. 2019. African Swine Fever Virus Armenia/07 Virulent Strain Controls Interferon Beta Production through the cGAS-STING Pathway. Journal of virology, 93, e02298-02218.

Gottipati K, Holthauzen L M, Ruggli N and Choi K H. 2016. Pestivirus Npro Directly Interacts with Interferon Regulatory Factor 3 Monomer and Dimer. Journal of virology, 90, 7740-7747.

Haluwana D K, Cha J W, Weerawardhana A, Gamage N, Subasinghe A, Jheong W and Lee J S. 2025. African swine fever virus L11L interferes with antiviral responses by targeting the IRF3 and PKR. Cellular and Molecular Life Sciences, 82, 425.

Hao S, Zheng X, Zhu Y, Yao Y, Li S, Xu Y and Feng W H. 2023. African swine fever virus QP383R dampens type I interferon production by promoting cGAS palmitoylation. Frontiers in Immunology, 14, 1186916.

Huang L, Xu W, Liu H, Xue M, Liu X, Zhang K, Hu L, Li J, Liu X, Xiang Z, Zheng J, Li C, Chen W, Bu Z, Xiong T and Weng C. 2021. African Swine Fever Virus pI215L Negatively Regulates cGAS-STING Signaling Pathway through Recruiting RNF138 to Inhibit K63-Linked Ubiquitination of TBK1. Journal of Immunology, 207, 2754-2769.

Iwasaki A and Medzhitov R. 2010. Regulation of adaptive immunity by the innate immune system. Science, 327, 291-295.

Jiao P, Ma J, Zhao Y, Jia X, Zhang H, Fan W, Jia X, Bai X, Zhao Y, Lu Y, Zhang H, Guo J, Pang G, Zhang K, Fang M, Li M, Liu W, Smith G L and Sun L. 2024. The nuclear localization signal of monkeypox virus protein P2 orthologue is critical for inhibition of IRF3-mediated innate immunity. Emerging Microbes and Infections, 13, 2372344.

Li D, Yang W, Li L, Li P, Ma Z, Zhang J, Qi X, Ren J, Ru Y, Niu Q, Liu Z, Liu X and Zheng H. 2021. African Swine Fever Virus MGF-505-7R Negatively Regulates cGAS-STING-Mediated Signaling Pathway. Journal of Immunology, 206, 1844-1857.

Li Z, Chen W, Qiu Z, Li Y, Fan J, Wu K, Li X, Zhao M, Ding H, Fan S and Chen J. 2022. African Swine Fever Virus: A Review. Life (Basel), 12, 103.

Lin C, Zhang C, Chen N, Meurens F, Zhu J and Zheng W. 2024. How Does African Swine Fever Virus Evade the cGAS-STING Pathway? Pathogens, 13, 957.

Lin R, Genin P, Mamane Y and Hiscott J. 2000. Selective DNA binding and association with the CREB binding protein coactivator contribute to differential activation of alpha/beta interferon genes by interferon regulatory factors 3 and 7. Molecular And Cellular Biology, 20, 6342-6353.

Lin R, Mamane Y and Hiscott J. 1999. Structural and functional analysis of interferon regulatory factor 3: localization of the transactivation and autoinhibitory domains. Mol Cell Biol, 19, 2465-2474.

Lin R, Mamane Y and Hiscott J. 1999. Structural and Functional Analysis of Interferon Regulatory Factor 3: Localization of the Transactivation and Autoinhibitory Domains. Molecular And Cellular Biology, 19, 2465-2474.

Liu H, Zhu Z, Feng T, Ma Z, Xue Q, Wu P, Li P, Li S, Yang F, Cao W, Xue Z, Chen H, Liu X and Zheng H. 2021. African Swine Fever Virus E120R Protein Inhibits Interferon Beta Production by Interacting with IRF3 To Block Its Activation. Journal of virology, 95, e0082421.

Liu S, Cai X, Wu J, Cong Q, Chen X, Li T, Du F, Ren J, Wu Y T, Grishin N V and Chen Z J. 2015. Phosphorylation of innate immune adaptor proteins MAVS, STING, and TRIF induces IRF3 activation. Science, 347, aaa2630.

Luteijn R D, van Terwisga S R, Ver Eecke J E, Onia L, Zaver S A, Woodward J J, Wubbolts R W, Raulet D H and van Kuppeveld F J M. 2024. The activation of the adaptor protein STING depends on its interactions with the phospholipid PI4P. Science Signaling, 17, eade3643.

Ranathunga L, Abesinghe S, Cha J W, Dodantenna N, Chathuranga K, Weerawardhana A, Haluwana D K, Gamage N and Lee J S. 2025. Inhibition of STING-mediated type I IFN signaling by African swine fever virus DP71L. Veterinary Research, 56, 27.

Song J X, Villagomes D, Zhao H and Zhu M. 2022. cGAS in nucleus: The link between immune response and DNA damage repair. Frontiers in Immunology, 13, 1076784.

Sun M, Yu S, Ge H, Wang T, Li Y, Zhou P, Pan L, Han Y, Yang Y, Sun Y, Li S, Li L F and Qiu H J. 2022. The A137R Protein of African Swine Fever Virus Inhibits Type I Interferon Production via the Autophagy-Mediated Lysosomal Degradation of TBK1. Journal of virology, 96, e0195721.

Wang G, Xie M, Wu W and Chen Z. 2021. Structures and Functional Diversities of ASFV Proteins. Viruses, 13, 2124.

Wu Q, Lei Y, Zuo Y, Zhang J, Guo F, Xu W, Xie T, Wang D, Peng G, Wang X, Chen H, Fu Z, Cao G and Dai J. 2023. Interactome between ASFV and host immune pathway proteins. mSystems, 8, e0047123.

Yin Q, Tian Y, Kabaleeswaran V, Jiang X, Tu D, Eck M J, Chen Z J and Wu H. 2012. Cyclic di-GMP sensing via the innate immune signaling protein STING. Molecular Cell, 46, 735-745.

Yu Z, Tong L, Ma C, Song H, Wang J, Chai L, Wang C, Wang M, Wang C, Yan R, Fu Y, Jia M, Zhao W and Zhao C. 2024. The UAF1-USP1 Deubiquitinase Complex Stabilizes cGAS and Facilitates Antiviral Responses. Journal of Immunology, 212, 295-301.

Zhao D, Liu R, Zhang X, Li F, Wang J, Zhang J, Liu X, Wang L, Zhang J, Wu X, Guan Y, Chen W, Wang X, He X and Bu Z. 2019. Replication and virulence in pigs of the first African swine fever virus isolated in China. Emerging Microbes and Infections, 8, 438-447.

Zhu L, Jin J, Wang T, Hu Y, Liu H, Gao T, Dong Q, Jin Y, Li P, Liu Z, Huang Y, Liu X and Cao C. 2024. Ebola virus sequesters IRF3 in viral inclusion bodies to evade host antiviral immunity. Elife, 12, RP88122.

[1]	Shuangxi Zhang, Xinlin Wei, Hejing Shen, Qinhu Wang, Yi Qiang, Langjun Cui, Hongxing Xu, Yuyan An, Meixiang Zhang. *Simultaneously enhancing plant growth and immunity through the application of engineered Bacillus subtilis* expressing a microbial pattern**[J]. >Journal of Integrative Agriculture, 2026, 25(5): 0-.
[2]	Jie Liu, Jie Zhang, Huijuan Yan, Tuyong Yi, Won Bo Shim, Zehua Zhou. **FvVam6 is associated with fungal development and fumonisin biosynthesis via vacuole morphology regulation in Fusarium verticillioides** [J]. >Journal of Integrative Agriculture, 2026, 25(5): 0-.
[3]	Saisai Wu, Shuqing Han, Jing Zhang, Guodong Cheng, Yali Wang, Kai Zhang, Mingming Han, Jianzhai Wu. Multi-scale keypoints detection and motion features extraction in dairy cows using ResNet101-ASPP network[J]. >Journal of Integrative Agriculture, 2026, 25(5): 0-.
[4]	Yijun Wang, Jinhao Han, Tenglong Zhang, Mengjia Sun, Hongyu Ren, Cunyao Bo, Yuqing Diao, Xin Ma, Hongwei Wang, Xiaoqian Wang. *Identification and fine mapping of a major QTL, qGPC4D, for grain protein content using wheat–Aegilops* tauschii introgression lines**[J]. >Journal of Integrative Agriculture, 2026, 25(5): 0-.
[5]	Zhenbang Zhu, Zhengqin Ye, Wenqiang Wang, Yanhua Li, Zhe Sun, Xiuling Yu, Kegong Tian, Xiangdong Li. A rescued virus from the infectious clone of a PRRSV NADC34-like strain exhibits high pathogenicity for nursery pigs[J]. >Journal of Integrative Agriculture, 2026, 25(5): 0-.
[6]	Xin Zhang, Jidong Zhang, Yunling Peng, Xun Yu, Lirong Lu, Yadong Liu, Yang Song, Dameng Yin, Shaogeng Zhao, Hongwu Wang, Xiuliang Jin, Jun Zheng. QTL mapping of maize plant height based on a population of doubled haploid lines using UAV LiDAR high-throughput phenotyping data[J]. >Journal of Integrative Agriculture, 2026, 25(5): 0-.
[7]	Chenyu Li, Zumuremu Tuerxun, Yang Yang, Xiaorong Li, Fengjiao Hui, Juan Li, Zhigang Liu, Guo Chen, Darun Cai, Hui Zhang, Xunji Chen, Shuangxia Jin, Bo Li. *Application of an endogenous pGhαGloA* promoter in CRISPR/Cas12a system for efficient genome editing to creat glandless cotton germplasm**[J]. >Journal of Integrative Agriculture, 2026, 25(5): 0-.
[8]	Mengyao Zhang, Hongli Jin, Cuicui Jiao, Yuanyuan Zhang, Yujie Bai, Zhiyuan Gong, Pei Huang, Haili Zhang, Yuanyuan Li, Hualei Wang. A candidate tick-borne encephalitis virus vaccine based on virus-like particles induces specific cellular and humoral immunity in mice[J]. >Journal of Integrative Agriculture, 2026, 25(5): 0-.
[9]	Mengsi Zhang, Mingming Yang, Xiaoxue Zhang, Shuying Li, Shuaiwu Wang, Alex Muremi Fulano, Yongting Meng, Xihui Shen, Lili Huang, Yao Wang. Two-Component Signaling System RegAB Represses Pseudomonas syringae pv. actinidiae T3SS by Directly Binding to the promoter of hrpRS[J]. >Journal of Integrative Agriculture, 2026, 25(5): 0-.
[10]	Jiyu Zhao, Xudong Sun, Yuqi Xue, Alam Sher, Jiayu Ran, Peng Liu, Bin Zhao, Baizhao Ren, Ningning Yu, Hao Ren, Jiwang Zhang. Optimizing nitrogen management for grain yield and nitrogen use efficiency in summer maize via coordinating the N supply–demand balance[J]. >Journal of Integrative Agriculture, 2026, 25(5): 0-.
[11]	Renxu Chang, Yuanyuan Chen, Xinyi Xu, Hongdou Jia, John Mauck, Juan J. Loor, Yehoshav A. Ben Meir, Qiushi Xu, Xudong Sun, Chuang Xu. Free fatty acids induce apoptosis in mammary epithelial cells from ketotic dairy cows via endoplasmic reticulum stress[J]. >Journal of Integrative Agriculture, 2026, 25(5): 0-.
[12]	Yeon Ju An, Min Young Kim, Sungup Kim, Jeongeun Lee, Sang Woo Kim, Jung In Kim, Eunyoung Oh, Heungsu Lee, Kwang-Soo Cho, Seung-Hyun Kim, Myoung Hee Lee, Eunsoo Lee. *QTL mapping and allele stacking for enhanced lignan content in sesame (Sesamum* indicum L.) using genotyping-by-sequencing**[J]. >Journal of Integrative Agriculture, 2026, 25(5): 0-.
[13]	Jingui Wei, Qiang Chai, Wen Yin, Yao Guo, Zhilong Fan, Falong Hu, Qiming Wang, Shoufa Mao. Mixed cropping green manure can simultaneously improve nutrition production and quality of spring wheat grain under reduced chemical nitrogen supply[J]. >Journal of Integrative Agriculture, 2026, 25(5): 0-.
[14]	Ying Liu, Jiangyao Fu, Haotian Chen, Yajun Zhang, Siyu Li, Kuanyu Zhu, Yunji Xu, Weilu Wang, Junfei Gu, Hao Zhang, Zhiqin Wang, Lijun Liu, Jianhua Zhang, Weiyang Zhang, Jianchang Yang. Cytokinins redistributing drives nitrogen remobilization from source to sink in wheat under moderate water limitation during grain filling[J]. >Journal of Integrative Agriculture, 2026, 25(5): 0-.
[15]	Qian Tang, Jianhong Ren, Xinru Zhang, Cai Wu, Yarong Zhang, Dahong Bian, Guangzhou Liu, Yanhong Cui, Xiong Du, Chuang Wang, Zhen Gao. Plant growth retardant increases nitrogen utilization efficiency and harvest index in maize by optimizing Plant Horizontal-Vertical Ratio and vascular bundles morphology[J]. >Journal of Integrative Agriculture, 2026, 25(5): 0-.

No Suggested Reading articles found!

Viewed

Full text

Abstract

Cited

Shared

Discussed

Cite this article:

About JIA

Editorial board

For authors