title: Isolation and characterization of cDNAs encoding Ars2 and Pasha homologues, two components of the RNA interference pathway in Litopenaeus vannamei.
abstract: The RNA interference (RNAi) is an evolutionarily conserved protective mechanism in eukaryotes against parasitic foreign nucleic acids. Previous studies demonstrated that the RNAi mechanism is important for shrimp antiviral immunity. Here, we report the identification and functional analysis of two key components of the shrimp RNAi activity: Litopenaeus vannamei arsenite resistance gene 2 (LvArs2) and partner of drosha (LvPasha). The full-length cDNA of LvArs2 was 3470 bp, including a 5' untranslated region (UTR) of 167 bp, a 3' UTR of 639 bp, and an open reading frame (ORF) of 2664 bp that encoded 887 amino acid residues with an estimated molecular mass of 102.5 kDa. The full-length cDNA of LvPasha was 2654 bp, including a 5' UTR of 99 bp, a 3' UTR of 560 bp, and an ORF of 1995 bp that encoded 664 amino acid residues with an estimated molecular mass of 74.2 kDa. Co-immunoprecipitation demonstrated that LvArs2 interacted with L. vannamei Dicer2 (LvDcr2) and LvPasha in Drosophila Schneider 2 (S2) cells, suggesting that LvArs2 may be involved in regulation of the miRNA/siRNA pathways in L.vannamei. Subcellular localization assays demonstrated both LvArs2 and LvPasha proteins mainly presented in the nucleus. After Poly(C-G) stimulation, the expression of LvArs2 was suppressed and expression of LvPasha was enhanced in shrimp gills. These results suggest that LvArs2 and LvPasha may participate in the defense against RNA viruses in crustacea.
authors.affiliation: MOE Key Laboratory of Aquatic Product Safety/State Key Laboratory for Biocontrol, School of Life Sciences, Sun Yat-sen University, 135 Xingang Road West, Guangzhou 510275, PR China.
authors.forename: Yi-Hong
authors.initials: YH
authors.lastname: Chen
authors.forename: Li
authors.initials: L
authors.lastname: Zhao
authors.forename: Xiao-Ting
authors.initials: XT
authors.lastname: Jia
authors.forename: Xiao-Yun
authors.initials: XY
authors.lastname: Li
authors.forename: Chao Zheng
authors.initials: CZ
authors.lastname: Li
authors.forename: Hui
authors.initials: H
authors.lastname: Yan
authors.forename: Shao-Ping
authors.initials: SP
authors.lastname: Weng
authors.forename: Jian-Guo
authors.initials: JG
authors.lastname: He
mesh_terms: D000276:Adjuvants, Immunologic / Q000494:pharmacology; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D018076:DNA, Complementary / Q000235:genetics*; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000187:drug effects; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D033561:Penaeidae / Q000145:classification / Q000235:genetics* / Q000276:immunology* / Q000821:virology; D010802:Phylogeny; D011066:Poly C / Q000494:pharmacology; D011068:Poly G / Q000494:pharmacology; D034622:RNA Interference; D016601:RNA-Binding Proteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D016415:Sequence Alignment
chemical_list: D000276:Adjuvants, Immunologic; D018076:DNA, Complementary; D009687:Nuclear Proteins; D016601:RNA-Binding Proteins; D011068:Poly G; C008961:poly G-poly C; D011066:Poly C