Identification, characterization, and function analysis of the Cactus gene from Litopenaeus vannamei

• Published in: PloS one Vol. 7; no. 11; p. e49711
• Main Authors: Li, Chaozheng, Chen, Yi-Xiao, Zhang, Shuang, Lü, Ling, Chen, Yi-Hong, Chai, Jiaoting, Weng, Shaoping, Chen, Yong-Gui, He, Jianguo, Xu, Xiaopeng
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 21.11.2012; Public Library of Science (PLoS)
• Subjects: Agriculture; Amino Acid Motifs; Amino Acid Sequence; Analysis; Animals; Ankyrins; Antimicrobial peptides; Bacteria; Biology; Chromatin; Cloning, Molecular; Computational Biology - methods; Crustaceans; Cytoplasm; DNA, Complementary - metabolism; DNA-Binding Proteins - genetics; Drosophila; Drosophila melanogaster; Drosophila Proteins - genetics; Feedback; Function analysis; Gene Expression Regulation; Genes; Genome; Health aspects; Homology; I-kappa B Proteins - metabolism; Immune response; Immunoprecipitation; Inhibitors; Innate immunity; Insects; Kinases; Lipopolysaccharides; Litopenaeus vannamei; Luciferase; Microscopy, Confocal - methods; Molecular Sequence Data; NF-κB protein; Nuclear transport; Overexpression; Penaeidae - genetics; Peptides; Phosphoproteins - genetics; Phylogeny; Polyinosinic:polycytidylic acid; Proteins; Sequence Homology, Amino Acid; Shellfish; Species Specificity; Transcription; Translocation; Viruses; Waterborne diseases; White spot syndrome
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0049711

The nuclear factor-kappa B (NF-κB) pathways play important roles in innate immune responses. IκB is the main cytoplasmic inhibitor of NF-κB. In this study, we identified the LvCactus gene from Litopenaeus vannamei, which is the first cloned IκB homologue in subphylum Crustacea. LvCactus contains six predicted ankyrin repeats, which show similarities to those of Cactus proteins from insects. LvCactus localizes in cytoplasm and interacts with LvDorsal, an L. vannamei homologue to Drosophila melanogaster Dorsal belonging to class II NF-κB family, to prevent its nuclear translocation. Contrary to that of LvDorsal, over-expression of LvCactus down-regulates the activities of shrimp antimicrobial peptides promoters, suggesting LvCactus is an inhibitor of LvDorsal. The promoter of LvCactus was predicted to contain five putative NF-κB binding motifs, among which four were proved to be bound by LvDorsal by chromatin immunoprecipitation assays. Dual-luciferase reporter assays also showed that transcription of LvCactus was promoted by LvDorsal but inhibited by LvCactus itself, indicating a feedback regulatory pathway between LvCactus and LvDorsal. Expression of LvCactus was up-regulated after Lipopolysaccharides, poly (I:C), Vibrio parahaemolyticus, and Staphylococcus aureus injections, suggesting an activation response of LvCactus to bacterial and immune stimulant challenges. Differently, the LvCactus expression levels obviously decreased during white spot syndrome virus (WSSV) infection, indicating the feedback regulatory pathway of LvCactus/LvDorsal could be modified by WSSV.