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凡纳滨对虾四种新型半胱天冬酶（Lvcaspase2 - 5）的鉴定及其通过双链RNA介导的基因沉默在对虾白斑综合征病毒感染中的作用

Characterization of four novel caspases from Litopenaeus vannamei (Lvcaspase2-5) and their role in WSSV infection through dsRNA-mediated gene silencing.

作者信息

Wang Pei-Hui, Wan Ding-Hui, Chen Yong-Gui, Weng Shao-Ping, Yu Xiao-Qiang, He Jian-Guo

机构信息

MOE Key Laboratory of Aquatic Product Safety/State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-Sen University, Guangzhou, People's Republic of China.

School of Marine Sciences, Sun Yat-Sen University, Guangzhou, People's Republic of China.

出版信息

PLoS One. 2013 Dec 23;8(12):e80418. doi: 10.1371/journal.pone.0080418. eCollection 2013.

DOI:10.1371/journal.pone.0080418
PMID:24376496
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3871164/
Abstract

Apoptosis plays an important role in white spot syndrome virus (WSSV) pathogenesis, and caspases are central players in apoptosis. Here, we cloned four novel caspases (Lvcaspase2-5) from the Pacific white shrimp Litopenaeus vannamei, and investigated their potential roles in WSSV replication using dsRNA-mediated gene silencing. Lvcaspase2-5 have the typical domain structure of caspase family proteins, with the conserved consensus motifs p20 and p10. Lvcaspase2 and Lvcaspase5 were highly expressed in muscle, while Lvcaspase3 was highly expressed in hemocytes and Lvcaspase4 was mainly expressed in intestine. Lvcaspase2-5 could also be upregulated by WSSV infection, and they showed different patterns in various tissues. When overexpressed in Drosophila S2 cells, Lvcaspase2-5 showed different cellular localizations. Using dsRNA-medicated gene silencing, the expression of Lvcaspase2, Lvcaspase3, and Lvcaspase5 were effectively knocked down. In Lvcaspase2-, Lvcaspase3- or Lvcaspase5-silenced L. vannamei, expression of WSSV VP28 gene was significantly enhanced, suggesting protective roles for Lvcaspase2, Lvcaspase3 and Lvcaspase5 in the host defense against WSSV infection.

摘要

细胞凋亡在白斑综合征病毒（WSSV）致病过程中发挥着重要作用，而半胱天冬酶是细胞凋亡的核心参与者。在此，我们从南美白对虾凡纳滨对虾中克隆了四种新型半胱天冬酶（Lvcaspase2 - 5），并使用双链RNA介导的基因沉默技术研究了它们在WSSV复制中的潜在作用。Lvcaspase2 - 5具有半胱天冬酶家族蛋白的典型结构域，带有保守的共有基序p20和p10。Lvcaspase2和Lvcaspase5在肌肉中高表达，而Lvcaspase3在血细胞中高表达，Lvcaspase4主要在肠道中表达。Lvcaspase2 - 5也可被WSSV感染上调，并且它们在不同组织中表现出不同模式。当在果蝇S2细胞中过表达时，Lvcaspase2 - 5表现出不同的细胞定位。使用双链RNA介导的基因沉默技术，Lvcaspase2、Lvcaspase3和Lvcaspase5的表达被有效敲低。在Lvcaspase2、Lvcaspase3或Lvcaspase5沉默的凡纳滨对虾中，WSSV VP28基因的表达显著增强，表明Lvcaspase2、Lvcaspase3和Lvcaspase5在宿主抵御WSSV感染中起保护作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6ccb/3871164/f18d4f26043b/pone.0080418.g009.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6ccb/3871164/cd257e38c72f/pone.0080418.g001.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6ccb/3871164/930293624344/pone.0080418.g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6ccb/3871164/8a0b39eaae1f/pone.0080418.g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6ccb/3871164/b520916e7a5f/pone.0080418.g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6ccb/3871164/d41580819317/pone.0080418.g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6ccb/3871164/9d79054f068b/pone.0080418.g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6ccb/3871164/f18d4f26043b/pone.0080418.g009.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6ccb/3871164/cd257e38c72f/pone.0080418.g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6ccb/3871164/2599fb00e149/pone.0080418.g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6ccb/3871164/e1ecdaaf5dea/pone.0080418.g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6ccb/3871164/930293624344/pone.0080418.g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6ccb/3871164/8a0b39eaae1f/pone.0080418.g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6ccb/3871164/b520916e7a5f/pone.0080418.g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6ccb/3871164/d41580819317/pone.0080418.g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6ccb/3871164/9d79054f068b/pone.0080418.g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6ccb/3871164/f18d4f26043b/pone.0080418.g009.jpg

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