title: β-Thymosins participate in antiviral immunity of red swamp crayfish (Procambarus clarkii).
abstract: β-Thymosins participate in numerous biological activities, including cell proliferation and differentiation, wound healing, and anti-inflammatory and antimicrobial activities. Many studies have investigated vertebrate β-thymosins, whereas few reports have focused on invertebrate β-thymosins. In this study, nine isoforms of β-thymosins (PcThy-1 to PcThy-8) were identified from the red swamp crayfish Procambarus clarkii. The isoforms contained different numbers of the thymosin β actin-binding motif. PcThy-1 contained one thymosin β actin-binding motif, whereas PcThy-8 contained eight motifs. Western blot analysis with anti-PcThy-4 antibody showed that three to six isoforms were present in one tissue, and PcThy-4, PcThy-5, PcThy-6, and PcThy-7 were the main isoforms in several tissues. Time course expression analysis of PcThys at the protein level showed that PcThy-4 was upregulated in hemocytes and gills after white spot syndrome virus (WSSV) challenge. PcThy-4, which contained four thymosin β actin-binding motifs, was selected for further research. Tissue distribution analysis by quantitative real-time PCR showed that PcThy-4 was present in tissues of the hemocytes, heart, hepatopancreas, gills, stomach, and intestine at the transcriptional level. Transcriptional expression profiles showed that PcThy-4 was upregulated after WSSV challenge. In vivo RNAi and protein injection assay results showed that PcThy-4 inhibited the replication of WSSV in crayfish and enhanced the survival rate after WSSV infection. Furthermore, PcThy-4 promoted hemocyte phagocytosis of WSSV. Overall, results suggested that PcThys protected crayfish from WSSV infection and played an important role in antiviral immune response.
authors.affiliation: Shandong Provincial Key Laboratory of Animal Cells and Developmental Biology, School of Life Sciences, Shandong University, Jinan, Shandong 250100, China.
authors.forename: Xiu-Zhen
authors.initials: XZ
authors.lastname: Shi
authors.affiliation: Shandong Provincial Key Laboratory of Animal Cells and Developmental Biology, School of Life Sciences, Shandong University, Jinan, Shandong 250100, China.
authors.forename: Li-Jie
authors.initials: LJ
authors.lastname: Shi
authors.affiliation: Shandong Provincial Key Laboratory of Animal Cells and Developmental Biology, School of Life Sciences, Shandong University, Jinan, Shandong 250100, China.
authors.forename: Yan-Ran
authors.initials: YR
authors.lastname: Zhao
authors.affiliation: Shandong Provincial Key Laboratory of Animal Cells and Developmental Biology, School of Life Sciences, Shandong University, Jinan, Shandong 250100, China.
authors.forename: Xiao-Fan
authors.initials: XF
authors.lastname: Zhao
authors.affiliation: Shandong Provincial Key Laboratory of Animal Cells and Developmental Biology, School of Life Sciences, Shandong University, Jinan, Shandong 250100, China. Electronic address: jxwang@sdu.edu.cn.
authors.forename: Jin-Xing
authors.initials: JX
authors.lastname: Wang
mesh_terms: D020816:Amino Acid Motifs / Q000235:genetics; D000818:Animals; D003400:Astacoidea / Q000276:immunology*; D004266:DNA Virus Infections / Q000276:immunology*; D020869:Gene Expression Profiling; D005880:Gills / Q000276:immunology; D006434:Hemocytes / Q000276:immunology*; D007109:Immunity / Q000235:genetics; D010587:Phagocytosis; D020033:Protein Isoforms / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics; D013947:Thymosin / Q000235:genetics / Q000378:metabolism*; D015854:Up-Regulation; D014779:Virus Replication / Q000235:genetics; D046848:White spot syndrome virus 1 / Q000502:physiology*
chemical_list: D020033:Protein Isoforms; D034741:RNA, Small Interfering; D013947:Thymosin
keywords: Antiviral immunity; Innate immunity; Procambarus clarkii; White spot syndrome virus; β-thymosin