甜菜夜蛾胱天蛋白酶-5可变剪切参与调控苜蓿丫纹夜蛾核多角体病毒侵染诱导的宿主细胞凋亡

doi:10.1016/j.jia.2025.03.001

Journal of Integrative Agriculture ›› 2025, Vol. 24 ›› Issue (12): 4715-4731.DOI: 10.1016/j.jia.2025.03.001

甜菜夜蛾胱天蛋白酶-5可变剪切参与调控苜蓿丫纹夜蛾核多角体病毒侵染诱导的宿主细胞凋亡

收稿日期:2024-10-31 修回日期:2025-03-25 接受日期:2024-12-23 出版日期:2025-12-20 发布日期:2025-03-04

Alternative splicing of Spodoptera exigua caspase-5 is involved in regulating host-cell apoptosis induced by AcMNPV infection

Qianlong Yu, Xinyu Wu, Minghui Wang, Jie Li, Guiling Zheng, Changyou Li^#

Shandong Engineering Research Center for Environment-Friendly Agricultural Pest Management, College of Plant Health and Medicine, Qingdao Agricultural University, Qingdao 266109, China

Received:2024-10-31 Revised:2025-03-25 Accepted:2024-12-23 Online:2025-12-20 Published:2025-03-04
About author:Qianlong Yu, Mobile: +86-13210022808, E-mail: qlyu@qau.edu.cn; #Correspondence Changyou Li, Tel: +86-532-58957462, E-mail: cyli@qau.edu.cn
Supported by:
The study was funded by the National Natural Science Foundation of China (32202393), the Natural Science Foundation of Shandong Province, China (ZR2021QC190), the Science and Technology Benefiting the People Demonstration Project of Qingdao, China (24-1-8-xdny-10-nsh) and the Qingdao Agricultural University High-level Talent Fund, China (663/1120101).

摘要/Abstract

摘要：

胱天蛋白酶（Caspase）是调控细胞凋亡的关键基因，其能够通过可变剪切形成不同的可变剪切体，进而调控细胞凋亡过程。鳞翅目昆虫Caspase也有可变剪切现象，然而它们形成的剪切体的功能尚不清楚。我们通过克隆甜菜夜蛾胱天蛋白酶-5（SeCaspase-5）基因，获得了四种具有不同基因序列和蛋白功能结构域的剪切体，分别命名为SeCaspase-5a、SeCaspase-5b、SeCaspase-5c和SeCaspase-5d。这些剪切体分别在甜菜夜蛾Se-3细胞中过表达表明，SeCaspase-5a具有促凋亡功能，而SeCaspase-5b、SeCaspase-5c和SeCaspase-5d则没有。半定量PCR分析显示，SeCaspase-5的不同剪切体在苜蓿丫纹夜蛾核多角体病毒（Autographa californica multiple nucleopolyhedrovirus, AcMNPV）感染期间的表达存在显著差异。进一步将SeCaspase-5的不同剪切体构建到AcMNPV 的基因组bacmid中并转染Se-3细胞，发现SeCaspase-5a促进了细胞凋亡并导致病毒的产量下降，而SeCaspase-5b、SeCaspase-5c和SeCaspase-5d没有促进细胞凋亡，反而增加了病毒的产量。此外，SeCaspase-5的不同剪切体之间的互作分析表明，SeCaspase-5a能够与其自身及SeCaspase-5b、SeCaspase-5c和SeCaspase-5d发生直接的蛋白互作。而且，将这些剪切体在Se-3细胞中共表达，表明SeCaspase-5b、SeCaspase-5c和SeCaspase-5d在一定程度上抑制了SeCaspase-5a的促凋亡功能，相比SeCaspase-5a单独表达导致凋亡细胞的百分比降低了约20%。这些结果表明，SeCaspase-5具有可变剪切现象，并且参与调控杆状病毒侵染诱导的细胞凋亡。这些发现加深了对鳞翅目昆虫胱天蛋白酶功能的理解，并为研究杆状病毒诱导宿主细胞凋亡的机制提供了新的见解。

Abstract:

Caspases, which play key roles in cell apoptosis, undergo alternative splicing to form different splicing variants that can regulate the apoptotic process. Lepidopteran insect caspases undergo alternative splicing, although the functions of their splicing variants are still unclear. The Spodoptera exigua caspase-5 (SeCaspase-5) gene was cloned and found to produce four different splicing variants with different gene sequences and protein functional domains, which were named SeCaspase-5a, SeCaspase-5b, SeCaspase-5c and SeCaspase-5d. Overexpression of these variants in S. exigua cells (Se-3) showed that SeCaspase-5a had a proapoptotic function, whereas SeCaspase-5b, SeCaspase-5c and SeCaspase-5d did not. Semi-qPCR analysis revealed that the expression of the SeCaspase-5 variants significantly differed during Autographa californica multiple nucleopolyhedrovirus (AcMNPV) infection. Furthermore, the SeCaspase-5 variants were constructed into the AcMNPV bacmid and transfected into Se-3 cells, which revealed that SeCaspase-5a promoted cell apoptosis and reduced virus production, whereas SeCaspase-5b, SeCaspase-5c and SeCaspase-5d did not promote cell apoptosis but instead increased virus production. Moreover, an analysis of the interactions between the SeCaspase-5 variants revealed that SeCaspase-5a directly interacted with SeCaspase-5b, SeCaspase-5c and SeCaspase-5d. Coexpression of these variants in Se-3 cells also revealed that SeCaspase-5b, SeCaspase-5c and SeCaspase-5d inhibited the proapoptotic function of SeCaspase-5a, resulting in a reduction in the percentage of apoptotic cells by about 20%. These results indicate that SeCaspase-5 undergoes alternative splicing and is involved in regulating the apoptosis induced by baculovirus infection. These findings increase our understanding of the functions of lepidopteran insect caspases and provide new insights into the mechanism of host-cell apoptosis induced by baculoviruses.

Key words: apoptosis , SeCaspase-5 , alternative splicing , AcMNPV , Spodoptera exigua

Qianlong Yu, Xinyu Wu, Minghui Wang, Jie Li, Guiling Zheng, Changyou Li. 甜菜夜蛾胱天蛋白酶-5可变剪切参与调控苜蓿丫纹夜蛾核多角体病毒侵染诱导的宿主细胞凋亡[J]. Journal of Integrative Agriculture, 2025, 24(12): 4715-4731.

Qianlong Yu, Xinyu Wu, Minghui Wang, Jie Li, Guiling Zheng, Changyou Li. Alternative splicing of Spodoptera exigua caspase-5 is involved in regulating host-cell apoptosis induced by AcMNPV infection[J]. Journal of Integrative Agriculture, 2025, 24(12): 4715-4731.

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甜菜夜蛾胱天蛋白酶-5可变剪切参与调控苜蓿丫纹夜蛾核多角体病毒侵染诱导的宿主细胞凋亡

Alternative splicing of Spodoptera exigua caspase-5 is involved in regulating host-cell apoptosis induced by AcMNPV infection

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