College of Fisheries, Key Lab of Freshwater Animal Breeding, Ministry of Agriculture, Key Lab of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education, Huazhong Agricultural University, Wuhan, 430070, PR China.
College of Fisheries, Key Lab of Freshwater Animal Breeding, Ministry of Agriculture, Key Lab of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education, Huazhong Agricultural University, Wuhan, 430070, PR China.
Fish Shellfish Immunol. 2020 Jul;102:82-91. doi: 10.1016/j.fsi.2020.04.020. Epub 2020 Apr 14.
Carbohydrate sulfotransferases 11 (chst11) is one of the enzymes that synthesize chondroitin sulfate (CS), which has extensive immune functions in vitro and plays a critical role in mediating the infection of host by pathogenic microorganisms. However, whether it has immune functions in crayfish is still poorly understood. In our previous study of transcriptome, chst11 was differentially expressed in susceptible individuals and resistant individuals of Procambarus clarkii after white spot syndrome virus (WSSV) injection. Thus, in this study, the sequence of chst11 was obtained from P. clarkii for the first time and analyzed, and the expression pattern of chst11 was investigated. Besides, the purified recombinant protein of chst11 effect in protection in WSSV infection was explored. The full length of chst11 was 1536 bp with an 831-bp open reading frame (ORF), which encoding 276 amino acids residues with a calculated molecular mass of 33.1 kDa. The chst11 contains a Sulfotransfer_2 domain, one N-glycosylation site and three O-glycosylation sites. Phylogenetic analysis results showed that chst11 had the highest similarity to Penaeus vannamei (79.93%). The expression pattern of chst11 in different tissues indicated that chst11 was expressed highest in gut, gill and hypodermis, lowest in testicular duct, periesophageal nerve and hemocytes. The chst11 had different expression patterns in different tissues when the crayfish was challenged by WSSV, Aeromonas hydrophila and CpG ODN. Recombinant chst11 protein significantly reduced the amount of WSSV copy number in hepatopancreas at 6 h and 12 h post injection compared to the control group injected with bovine serum albumin (BSA). It was found that chst11 protein enhanced the expression of peroxinectin, proPO in hepatopancreas and midgut and the C-type lectin (ctl) in hemocytes and hepatopancreas. Intramuscularly injection of juvenile crayfish with chst11 protein decreased 60% mortality compared to the control group with BSA. This study is the first report on the antiviral function of chst11 in the immune system of crustacean.
碳水化合物磺基转移酶 11（chst11）是合成软骨素硫酸盐（CS）的酶之一，CS 在体外具有广泛的免疫功能，在介导宿主感染致病微生物方面起着关键作用。然而，它在小龙虾中是否具有免疫功能仍知之甚少。在我们之前对转录组的研究中，chst11 在白斑综合征病毒（WSSV）注射后易感性个体和抗性个体的 Procambarus clarkii 中差异表达。因此，在这项研究中，首次从 P. clarkii 中获得并分析了 chst11 的序列，并研究了 chst11 的表达模式。此外，还探索了 chst11 纯化重组蛋白在 WSSV 感染中的保护作用。chst11 的全长为 1536bp，含有 831bp 的开放阅读框（ORF），编码 276 个氨基酸残基，计算分子量为 33.1kDa。chst11 包含一个 Sulfotransfer_2 结构域、一个 N-糖基化位点和三个 O-糖基化位点。系统进化分析结果表明，chst11 与凡纳滨对虾（Penaeus vannamei）的相似度最高（79.93%）。chst11 在不同组织中的表达模式表明，chst11 在肠道、鳃和真皮中表达最高，在精巢管、围食管神经和血淋巴中表达最低。当小龙虾受到 WSSV、嗜水气单胞菌和 CpG ODN 挑战时，chst11 在不同组织中的表达模式不同。与注射牛血清白蛋白（BSA）的对照组相比，重组 chst11 蛋白在注射后 6 小时和 12 小时显著降低了肝胰腺中 WSSV 拷贝数的量。结果发现，chst11 蛋白增强了肝胰腺和中肠中的过氧化物酶、原 PO 和血淋巴和肝胰腺中的 C 型凝集素（ctl）的表达。与注射 BSA 的对照组相比，肌肉内注射 chst11 蛋白可使幼虾的死亡率降低 60%。本研究首次报道了 chst11 在甲壳动物免疫系统中的抗病毒功能。
