热休克蛋白27对水泡性口炎病毒体外增殖的调控作用

doi:10.11843/j.issn.0366-6964.2024.06.025

畜牧兽医学报 ›› 2024, Vol. 55 ›› Issue (6): 2540-2549.doi: 10.11843/j.issn.0366-6964.2024.06.025

热休克蛋白27对水泡性口炎病毒体外增殖的调控作用

李殿玉¹^,²(), 莫荣纤¹^,², 赵旭², 高铭², 李洪珊¹^,², 白辉盛², 马瑞仙¹^,², 李向茸¹^,³^,*(), 冯若飞¹^,³^,*()

1. 西北民族大学生物医学研究中心，生物工程与技术国家民委重点实验室，兰州 730030
2. 西北民族大学，生命科学与工程学院，兰州 730030
3. 西北民族大学生物医学研究中心，细胞基质疫苗关键技术与产业化教育部工程研究中心，兰州 730030

收稿日期:2023-06-26 出版日期:2024-06-23 发布日期:2024-06-28
通讯作者: 李向茸,冯若飞 E-mail:994398211@qq.com;lixiangrong@xbmu.edu.cn;fengruofei@xbmu.edu.cn
作者简介:李殿玉(1997-)，男，天津人，硕士生，主要从事病原生物学及基因工程研究，E-mail: 994398211@qq.com；Tel：0931-2928315
基金资助:
西北民族大学中央高校基本科研业务费资金项目(31920230162);国家自然科学基金(32260037);西北民族大学国家级大学生创新创业训练计划项目(202210742022);西北民族大学大学生创新训练项目(X202310742266)

Regulation of Heat Shock Protein 27 on the Proliferation of Vesicular Stomatitis Virus in vitro

Dianyu LI¹^,²(), Rongqian MO¹^,², Xu ZHAO², Ming GAO², Hongshan LI¹^,², Huisheng BAI², Ruixian MA¹^,², Xiangrong LI¹^,³^,*(), Ruofei FENG¹^,³^,*()

1. Key Laboratory of Biotechnology and Bioengineering of State Ethnic Affairs Commission, Biomedical Research Center, Northwest Minzu University, Lanzhou 730030, China
2. Life Science and Engineering College of Northwest Minzu University, Lanzhou 730030, China
3. Engineering Research Center of Key Technology and Industrialization of Cell-based Vaccine, Ministry of Education, Biomedical Research Center, Northwest Minzu University, Lanzhou 730030, China

Received:2023-06-26 Online:2024-06-23 Published:2024-06-28
Contact: Xiangrong LI, Ruofei FENG E-mail:994398211@qq.com;lixiangrong@xbmu.edu.cn;fengruofei@xbmu.edu.cn

摘要/Abstract

摘要：

本研究旨在探究热休克蛋白27(heat shock protein 27, HSP27)对水泡性口炎病毒(vesicular stomatitis virus, VSV)体外增殖及VSV感染介导的维甲酸诱导基因I样受体家族(retinoid acid-inducible gene-I-like receptor, RLR)信号通路的作用及机制。利用实时荧光定量PCR、免疫印迹、免疫共沉淀、病毒滴度测定及免疫荧光等方法，首先检测VSV感染对HSP27 mRNA和蛋白表达的影响，进而验证过表达和干扰HSP27对VSV增殖的影响，进一步探究HSP27对VSV感染介导的RLR信号通路的影响，最后分析HSP27与RLR信号通路靶分子的相互作用及共定位情况。结果表明，VSV感染可促使HSP27基因转录和蛋白表达上调，稳定过表达和瞬时过表达HSP27均能显著抑制VSV的体外增殖；干扰宿主细胞中HSP27表达可促进VSV增殖。HSP27能够增强VSV及维甲酸诱导基因I蛋白(retinoic acid-inducible gene I protein, RIG-I)介导的IFN-β的产生及RIG-I的蛋白表达，而且HSP27与RIG-I相互作用并共定位于细胞质中。本研究揭示了HSP27靶向RIG-I上调其表达，增强RLR信号通路的转导，进而负调控VSV体外增殖，为深入揭示宿主因子HSP27在病毒感染中的作用机制奠定基础。

关键词: 热休克蛋白27, 水泡性口炎病毒, RLR信号通路, 维甲酸诱导基因I蛋白

Abstract:

This study was conducted to investigate the effect and mechanism of heat shock protein 27 (HSP27) on the proliferation of vesicular stomatitis virus (VSV) in vitro and the retinoid acid-inducible gene-I-like receptor (RLR) signaling pathway mediated by VSV infection. Real-time quantitative PCR, Western blotting, Co-immunoprecipitation, TCID₅₀ assay, and immunofluorescence methods were employed to reveal the effects of VSV infection on the mRNA and protein expression of HSP27, and then the effects of overexpression and interference with HSP27 on the proliferation of VSV, and further explore the effects of HSP27 on the RLR signaling pathway mediated by VSV infection. Finally, the interaction and co-localization between HSP27 and the target molecules in RLR signaling pathway were analyzed. The results showed that VSV infection could increase the gene transcription and protein expression of HSP27, both stable and transient overexpression of HSP27 significantly inhibited the proliferation of VSV in vitro. Interfering with HSP27 expression in host cells promoted the proliferation of VSV. Further studies showed that HSP27 enhanced both VSV and retinoic acid-inducible gene I protein (RIG-I) mediated IFN-β production and the protein expression of RIG-I, and that HSP27 interacted with RIG-I and co-localized in the cytoplasm. This study for the revealed the molecular mechanism by which HSP27 targets RIG-I to upregulate its expression, enhance the transduction of RLR signaling pathways, and then negatively regulate the proliferation of VSV in vitro, laying a foundation for further revealing the mechanism of host factor HSP27 in viral infection.

Key words: heat shock protein 27, vesicular stomatitis virus, RLR signaling pathway, RIG-I

中图分类号:

S852.659.5

李殿玉, 莫荣纤, 赵旭, 高铭, 李洪珊, 白辉盛, 马瑞仙, 李向茸, 冯若飞. 热休克蛋白27对水泡性口炎病毒体外增殖的调控作用[J]. 畜牧兽医学报, 2024, 55(6): 2540-2549.

Dianyu LI, Rongqian MO, Xu ZHAO, Ming GAO, Hongshan LI, Huisheng BAI, Ruixian MA, Xiangrong LI, Ruofei FENG. Regulation of Heat Shock Protein 27 on the Proliferation of Vesicular Stomatitis Virus in vitro[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(6): 2540-2549.

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引物名称 Names	核苷酸序列(5′→3′) Nucleotide sequences
Homo-HSP27-qF	CTGACGGTCAAGACCAAGGATG
Homo-HSP27-qR	GTGTATTTCCGCGTGAAGCACC
Homo-GAPDH-qF	GTCTCCTCTGACTTCAACAGCG
Homo-GAPDH-qR	ACCACCCTGTTGCTGTAGCCAA
Homo-IFN-β-qF	TTGTTGAGAACCTCCTGGCT
Homo-IFN-β-qR	TGACTATGGTCCAGGCACAG

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热休克蛋白27对水泡性口炎病毒体外增殖的调控作用

Regulation of Heat Shock Protein 27 on the Proliferation of Vesicular Stomatitis Virus in vitro

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