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

• Published in: PloS one Vol. 8; no. 12; p. e80418
• Main Authors: Wang, Pei-Hui, Wan, Ding-Hui, Chen, Yong-Gui, Weng, Shao-Ping, Yu, Xiao-Qiang, He, Jian-Guo
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 23.12.2013; Public Library of Science (PLoS)
• Subjects: Animals; Apoptosis; Caspase; Caspases - genetics; Caspases - metabolism; Cell Line; Cloning; Cloning, Molecular; Double-stranded RNA; Drosophila; Fruit flies; Gene expression; Gene Expression Profiling; Gene Knockdown Techniques; Gene Silencing; Genes; Genetic engineering; Health aspects; Hemocytes; Infection; Infections; Intestine; Laboratories; Life sciences; Litopenaeus vannamei; Muscles; Pathogenesis; Penaeidae - enzymology; Penaeidae - virology; Penaeus monodon; Peptides; Phylogeny; Product safety; Protein Transport; Proteins; RNA, Double-Stranded - metabolism; Sequence Analysis, DNA; Shellfish; Subcellular Fractions - metabolism; Time Factors; Tissues; Tumor necrosis factor-TNF; Vibrio alginolyticus; Viral infections; Virus Replication; Viruses; White spot syndrome; White spot syndrome virus 1 - physiology
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0080418

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.