Autophagy induced by Vip3Aa has a pro-survival role in Spodoptera frugiperda Sf9 cells.
Autophagy induced by Vip3Aa has a pro-survival role in Spodoptera frugiperda Sf9 cells.
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DOI：
10.1080/21505594.2021.1878747
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发表时间：
2021-12
期刊：
影响因子：
5.2
通讯作者：
Cai J
中科院分区：
文献类型：
--
作者：
Hou X;Han L;An B;Cai J
Vip3Aa是一种杀虫蛋白，可以有效防治某些鳞翅目害虫，在生物防治中得到广泛应用。然而，Vip3Aa 的作用机制尚不清楚。在本研究中，我们发现Vip3Aa可以引起Sf9细胞中的自噬，这通过GFP-Atg8斑点数量的增加、自噬空泡的出现以及Atg8-II蛋白水平的升高得到证实。此外，我们发现 AMPK-mTOR-ULK1 通路参与 Vip3Aa 诱导的自噬，这可能与 Vip3Aa 处理的细胞中 ATP 稳态的破坏有关。 p62 水平升高和 GFP-RFP-Atg8 黄色荧光点数量增加表明，Sf9 细胞中的自噬在 Vip3Aa 处理后 24 小时受到抑制。随着Vip3Aa处理时间的延长，这种抑制变得更加严重并导致自噬体积累。 ATG5 的基因敲除或自噬抑制剂 3-MA 的使用进一步增加了 Sf9 细胞对 Vip3Aa 的敏感性。 ATG5 的过度表达降低了 Vip3Aa 处理细胞的细胞死亡率。综上所述，结果表明，Vip3Aa诱导的自噬具有促生存作用，这可能与昆虫抗性的发展有关。
Vip3Aa is an insecticidal protein that can effectively control certain lepidopteran pests and has been used widely in biological control. However, the mechanism of action of Vip3Aa is unclear. In the present study, we showed that Vip3Aa could cause autophagy in Sf9 cells, which was confirmed by the increased numbers of GFP-Atg8 puncta, the appearance of autophagic vacuoles, and an elevated Atg8-II protein level. Moreover, we found that the AMPK-mTOR-ULK1 pathway is involved in Vip3Aa-induced autophagy, which might be associated with the destruction of ATP homeostasis in Vip3Aa-treated cells. Both the elevated p62 level and the increased numbers of GFP-RFP-Atg8 yellow fluorescent spots demonstrated that autophagy in Sf9 cells was inhibited at 24 h after Vip3Aa treatment. With the prolongation of Vip3Aa treatment time, this inhibition became more serious and led to autophagosome accumulation. Genetic knockdown of ATG5 or the use of the autophagy inhibitor 3-MA further increased the sensitivity of Sf9 cells to Vip3Aa. Overexpression of ATG5 reduced the cell mortality of Vip3Aa-treated cells. In summary, the results revealed that autophagy induced by Vip3Aa has a pro-survival role, which might be related to the development of insect resistance.
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