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Caspase-8 in Labeo rohita is evolutionary conserved and is activated in Aeromonas hydrophila and Edwardsiella tarda infection and rhabdovirus vaccination
The Journal of Basic and Applied Zoology Pub Date : 2020-04-15 , DOI: 10.1186/s41936-020-00150-6
Mrinal Samanta , Alok Kumar Giri , Mahismita Paichha , Pushpa Choudhary , Ashis Saha

Caspase-8, a member of the family of conserved cysteine proteases, plays a crucial role in the initiation phase of the apoptotic death-signaling cascade and thereby attracts interest for its study across the animal species including fish. In India, rohu (Labeo rohita) is an important freshwater fish species; thus, this study on caspase-8 was undertaken to investigate its role during pathogenic invasion. The complete cDNA sequence of Labeo rohita caspase-8 (Lrcasp8) consisted of 1746 bp nucleotides (nt) having an ORF of 1440 nt encoding a polypeptide of 480 amino acid (aa) residues with the molecular mass of ∼ 54.8 kDa. Structurally, Lrcasp8 comprised two DED domains (DED11-77aa and DED297-174aa) and one CASc domain230-476aa. Within the CASc domain, various putative motifs, viz., a large subunit (p20237-360aa), a small subunit (p10389-474aa), and a penta-peptide (QACQG354-358aa) active site, were identified. The secondary structure of Lrcasp8 protein comprised seventeen α-helices, eleven β-strands, and twenty-nine coils. Phylogenetically, it is closely related to common carp caspase-8 and exhibits significant (p < 0.05) similarity (88.3%) and identity (78.7 %) in their amino acid sequence. The tissue-specific expression of Lrcasp8 has been analyzed by quantitative real-time PCR assay, and it revealed the highest expression (~ 23-fold) in the blood and lowest in the spleen. In Aeromonas hydrophila and Edwardsiella tarda infection and rhabdovirus vaccination, caspase-8 gene expression in rohu fingerlings was significantly (p <0.05) induced in various organs/tissues. Infection of the Labeo rohita gill cells with A. hydrophila resulted in apoptosis and cell death with the induction of caspase-8 gene expression. This is the first report on the identification and structural characterization of caspase-8 cDNA and predicted protein and the analysis of caspase-8 gene expression in Labeo rohita following Aeromonas hydrophila and Edwardsiella tarda infections and rhabdovirus vaccinations. The data in this article together suggest the critical role of caspase-8 during infection and apoptosis in Labeo rohita.

中文翻译:

Labeo rohita 中的 Caspase-8 在进化上是保守的,并且在嗜水气单胞菌和迟发性爱德华氏菌感染和弹状病毒疫苗接种中被激活

Caspase-8 是保守的半胱氨酸蛋白酶家族的成员,在凋亡死亡信号级联的起始阶段起着至关重要的作用,因此引起了对包括鱼类在内的动物物种研究的兴趣。在印度,罗湖鱼(Labeo rohita)是一种重要的淡水鱼类;因此,对 caspase-8 进行了这项研究,以研究其在病原体入侵过程中的作用。Labeo rohita caspase-8 (Lrcasp8) 的完整 cDNA 序列由 1746 bp 核苷酸 (nt) 组成,ORF 为 1440 nt,编码 480 个氨基酸 (aa) 残基的多肽,分子量约为 54.8 kDa。在结构上,Lrcasp8 包含两个 DED 域(DED11-77aa 和 DED297-174aa)和一个 CASc 域 230-476aa。在 CASc 域内,各种假定的基序,即大亚基 (p20237-360aa)、小亚基 (p10389-474aa)、和五肽 (QACQG354-358aa) 活性位点被鉴定出来。Lrcasp8 蛋白的二级结构包括 17 个 α-螺旋、11 个 β-链和 29 个螺旋。在系统发育上,它与鲤鱼 caspase-8 密切相关,并且在它们的氨基酸序列中表现出显着的 (p < 0.05) 相似性 (88.3%) 和同一性 (78.7%)。Lrcasp8 的组织特异性表达已通过定量实时 PCR 测定进行分析,它揭示了血液中的最高表达(~ 23 倍)和脾脏中的最低表达。在嗜水气单胞菌和迟发性爱德华氏菌感染和弹状病毒疫苗接种中,罗狐鱼种中的 caspase-8 基因表达在各种器官/组织中被显着诱导(p <0.05)。用 A. 感染 Labeo rohita 鳃细胞。Hydrophila 通过诱导 caspase-8 基因表达导致细胞凋亡和细胞死亡。这是关于 caspase-8 cDNA 和预测蛋白的鉴定和结构表征以及在嗜水气单胞菌和迟发性爱德华氏菌感染和弹状病毒疫苗接种后 Labeo rohita 中 caspase-8 基因表达分析的第一份报告。本文中的数据共同表明 caspase-8 在 Labeo rohita 感染和细胞凋亡过程中的关键作用。
更新日期:2020-04-15
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