Molecular characterization and expression of the autophagy-related gene Atg14 in WSSV-infected Procambarus clarkii

文献类型： 外文期刊

第一作者： Zhu, Mengru

作者： Zhu, Mengru;Zhan, Ming;Xi, Changjun;Gong, Jie;Shen, Huaishun;Zhu, Mengru;Zhan, Ming;Xi, Changjun;Gong, Jie;Shen, Huaishun

作者机构：

关键词： Atg14; Procambarus clarkii; WSSV; Autophagy; RNAi

期刊名称：FISH & SHELLFISH IMMUNOLOGY （ 影响因子：4.622； 五年影响因子：4.799 ）

ISSN： 1050-4648

年卷期： 2022 年 125 卷

页码：

收录情况： SCI

摘要： Atg14 (autophagy-related gene 14), also known as Atg14L or Barkor (Beclin-1 associated autophagy-related key regulator), plays an important role in a variety of biological processes including immunity, development, tumor inhibition, longevity, and protection against some cardiac and neurodegenerative diseases. However, very few studies have characterized Atg14 expression in invertebrates, particularly crustaceans. Here, a novel Atg14 gene from Procambarus clarkii (named PcAtg14) was characterized via RACE technology. Bioinformatics analysis showed that the total length of the PcAtg14 gene sequence was 2,880 bp, and it was predicted to encode 488 amino acids. The results of homology comparison showed that PcAtg14 exhibited the highest homology with crustacean the American lobster (Homarus americanus). Quantitative real-time PCR expression analysis showed that PcAtg14 was expressed in all tissues of P. clarkii, with the hepatopancreas having the highest expression and the eyestalk exhibiting the lowest expression. Upon white spot syndrome virus (WSSV) infection, the relative expression of PcAtg14 in the hepatopancreas, muscle, hemocyte, gill, heart and epidermis were significantly up regulated at different time periods. After PcAtg14 gene silencing via RNA interference (RNAi), the proliferation of WSSV in P. clarkii was significantly inhibited, which coincided with a significant increase in P. clarkii mortality and an increase in the expression of autophagy-related genes (ATGs). Transmission electron microscopy analysis demonstrated an increase in the number of autophagosomes in the hepatopancreas of the PcAtg14 gene silencing group compared to the control group after WSSV infection. Collectively, these results indicated that PcAtg14 suppressed autophagy by reduce the fusion of autophagosomes and lysosomes, thereby promoting WSSV replication in P. clarkii. The findings here therefore provide novel insights into the immune mechanisms through which P. clarkii responds to WSSV infection.

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