Function investigation of p11.5 in ASFV infection

Dan Yin; Bin Shi; Renhao Geng; Yingnan Liu; Lang Gong; Hongxia Shao; Kun Qian; Hongjun Chen; Aijian Qin

doi:10.1016/j.virs.2024.05.007

June 2024

Dan Yin, Bin Shi, Renhao Geng, Yingnan Liu, Lang Gong, Hongxia Shao, Kun Qian, Hongjun Chen and Aijian Qin. Function investigation of p11.5 in ASFV infection[J]. Virologica Sinica, 2024, 39(3): 469-477. doi: 10.1016/j.virs.2024.05.007

Citation: Dan Yin, Bin Shi, Renhao Geng, Yingnan Liu, Lang Gong, Hongxia Shao, Kun Qian, Hongjun Chen, Aijian Qin. Function investigation of p11.5 in ASFV infection .VIROLOGICA SINICA, 2024, 39(3) : 469-477. http://dx.doi.org/10.1016/j.virs.2024.05.007

非洲猪瘟病毒p11.5蛋白在病毒感染中的作用

尹丹 ^a,b,c ,
石彬 ^a,b,c ,
耿仁浩 ^a,b,c ,
刘英楠 ^d ,
龚浪 ^e ,
邵红霞 ^a,b,c ,
钱琨 ^a,b,c,, ,
陈鸿军 ^d,, ,
秦爱建 ^a,b,c,,

cstr: 32224.14.j.virs.2024.05.007

a. 扬州大学教育部禽类预防医学省部共建重点实验室江苏省动物重要疫病与人畜共患病防控协同控制中心;
b. Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Jiangsu 225009, China;
c. 教育部农业与农产品安全国际合作联合实验室;
d. 中国农业科学院上海兽医研究所;
e. 华南农业大学

通讯作者： 钱琨, qiankun@yzu.edu.cn; 陈鸿军, vetchj@shvri.ac.cn; 秦爱建, aijian@yzu.edu.cn
收稿日期： 2023-11-29
录用日期： 2024-05-17

摘要

病毒复制依赖于蛋白质之间复杂的相互作用。就非洲猪瘟病毒而言，目前仅有少数蛋白质之间的相互作用被鉴定。因此，本研究利用免疫共沉淀和液相色谱-质谱技术证明非洲猪瘟病毒 p72 蛋白与A137R基因编码的 p11.5 蛋白之间存在互作关系。研究发现，p72 蛋白的1-216位氨基酸与 p11.5 蛋白的1-68位氨基酸是两者互作的关键结构域。为了进一步评估p11.5蛋白在病毒感染中的作用，本研究构建了一种缺失A137R基因的重组病毒（ASFVGZΔA137R），与亲本病毒ASFVGZ相比，ASFVGZΔA137R的病毒滴度显著降低，但ASFVGZΔA137R感染细胞后子代病毒中的基因组拷贝数与病毒滴度的比值更高，据此推测感染ASFVGZΔA137R的细胞可能产生了大量非感染性的病毒颗粒。同时我们发现 p11.5 不影响病毒与细胞的结合和内吞作用。本研究首次证明了 p72 和 p11.5 之间的相互作用，加深了我们对病毒蛋白之间复杂相互作用的理解，为进一步研究非洲猪瘟病毒感染的致病机制提供了参考。
- 非洲猪瘟病毒
- , p72 蛋白
- , p11.5 蛋白
- , 蛋白相互作用

Function investigation of p11.5 in ASFV infection

Dan Yin ^a,b,c ,
Bin Shi ^a,b,c ,
Renhao Geng ^a,b,c ,
Yingnan Liu ^d ,
Lang Gong ^e ,
Hongxia Shao ^a,b,c ,
Kun Qian ^a,b,c,, ,
Hongjun Chen ^d,, ,
Aijian Qin ^a,b,c,,

a. Ministry of Education Key Laboratory for Avian Preventive Medicine, Yangzhou University, Jiangsu 225009, China;
b. Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Jiangsu 225009, China;
c. Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Jiangsu 225009, China;
d. Shanghai Veterinary Research Institute, CAAS, Shanghai, 200241, China;
e. South China Agricultural University, Guangzhou 510642, China

Corresponding author: Kun Qian, qiankun@yzu.edu.cn Hongjun Chen, vetchj@shvri.ac.cn Aijian Qin, aijian@yzu.edu.cn
Received Date: 29 November 2023
Accepted Date: 17 May 2024

Abstract

Virus replication relies on complex interactions between viral proteins. In the case of African swine fever virus (ASFV), only a few such interactions have been identified so far. In this study, we demonstrate that ASFV protein p72 interacts with p11.5 using co-immunoprecipitation and liquid chromatography-mass spectrometry (LC-MS). It was found that protein p72 interacts specifically with p11.5 at sites amino acids (aa) 1-216 of p72 and aa 1-68 of p11.5. To assess the importance of p11.5 in ASFV infection, we developed a recombinant virus (ASFVGZΔA137R) by deleting the A137R gene from the ASFVGZ genome. Compared with ASFVGZ, the infectious progeny virus titers of ASFVGZΔA137R were reduced by approximately 1.0 logs. In addition, we demonstrated that the growth defect was partially attributable to a higher genome copies-to-infectious virus titer ratios produced in ASFVGZΔA137R-infected MA104 cells than in those infected with ASFVGZ. This finding suggests that MA104 cells infected with ASFVGZΔA137R may generate larger quantities of noninfectious particles. Importantly, we found that p11.5 did not affect virus-cell binding or endocytosis. Collectively, we show for the first time the interaction between ASFV p72 and p11.5. Our results effectively provide the relevant information of the p11.5 protein. These results extend our understanding of complex interactions between viral proteins, paving the way for further studies of the potential mechanisms and pathogenesis of ASFV infection.
- African swine fever virus
- , p72 protein
- , p11.5 protein
- , Protein interactions

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Electronic Supplementary Material

10.1016j.virs.2024.05.007-ESM.docx