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Functional identification of Bombyx mori Atg13 in autophagy.
Archives of Insect Biochemistry and Physiology ( IF 2.2 ) Pub Date : 2020-06-09 , DOI: 10.1002/arch.21718
Shuyan Li 1 , Jing Xu 1 , Xianying Xu 1 , Weiyuan Ma 1 , Ling Tian 1 , Kang Li 2
Affiliation  

The autophagy process involves a series of autophagy‐related (Atg) proteins, which are conserved in eukaryotes. ULK1/Atg1‐ATG13/Atg13 is the core protein complex for autophagy initiation in response to nutrient and hormone signaling. However, how Atg13 is regulated to participate in autophagy is unclear in insects. Here in Bombyx mori, the variation of BmAtg13 was correlated with autophagy induced by steroid hormone 20‐hydroxyecdysone (20E) or starvation. Developmental profiles from feeding to prepupal stage revealed that there were two bands of BmAtg13 protein detected by western blot analysis, therein the upper band was intensively decreased, while the lower band was significantly increased which was in accordance with its mRNA variation; and immunofluorescent staining indicated that BmAtg13 was nucleocytoplasmic translocated during larval‐pupal metamorphosis when autophagy was dramatically induced. BmAtg13 knockdown and overexpression both inhibits autophagy. Besides, 20E treatment‐induced BmAtg13 gene expression, while blocking 20E signaling transduction by knockdown of BmUsp reduced both gene expression and protein level of BmAtg13. These results reveal that BmAtg13 is required for 20E‐ and starvation‐induced autophagy in B. mori, which provides the foundation for further related studies.

中文翻译:

自噬中家蚕Atg13的功能鉴定。

自噬过程涉及一系列自噬相关(Atg)蛋白质,这些蛋白质在真核生物中是保守的。ULK1 / Atg1-ATG13 / Atg13是响应营养和激素信号而自噬引发的核心蛋白复合物。但是,尚不清楚昆虫如何调控Atg13参与自噬。这里在家蚕,BmAtg13的变化与类固醇激素20-羟基蜕皮激素(20E)或饥饿引起的自噬相关。从进食到成骨前阶段的发育概况显示,通过蛋白质印迹分析检测到BmAtg13蛋白有两个条带,其中上条带被强烈减少,而下条带则被显着增加,这与其mRNA的变化一致。免疫荧光染色表明,当自噬被强烈诱导时,BmAtg13在幼虫-pal变态期间是核质转移的。BmAtg13敲低和过度表达都抑制自噬。此外,20E治疗诱导BmAtg13基因表达,同时通过敲低BmUsp阻断20E信号转导。降低BmAtg13的基因表达和蛋白质水平。这些结果表明,BmAtg13需要20F-和饥饿诱导自噬家蚕,这提供了进一步的相关研究奠定了基础。
更新日期:2020-06-09
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