Department of Biology and Guangdong Provincial Key Laboratory of Marine Biotechnology, Shantou University, Shantou 515063, China.
Department of Biology and Guangdong Provincial Key Laboratory of Marine Biotechnology, Shantou University, Shantou 515063, China.
Fish Shellfish Immunol. 2018 Mar;74:491-500. doi: 10.1016/j.fsi.2018.01.016. Epub 2018 Jan 31.
The Pim1 serine/threonine kinase is associated with multiple cellular functions including proliferation, survival, differentiation, apoptosis, tumorigenesis, immune regulation and inflammation in vertebrates. However, little is known about the role of Pim1 in invertebrate immunity. In this study, we identified and characterized for the first time, a Pim1 (LvPim1) gene in Litopenaeus vannamei, with a full-length cDNA of 2352 bp and a 1119 bp open reading frame (ORF) encoding a putative protein of 372 amino acids, which contains a typical serine/threonine kinase domain. Sequence and phylogenetic analysis revealed that LvPim1 shared a close evolutionary relationship with Pim1 from vertebrates. Real-time qPCR analysis showed that LvPim1 was widely expressed in all tissues tested; with its transcript level induced in hepatopancreas and hemocytes upon challenge with Vibrio parahaemolyticus, Streptoccocus iniae, lipopolysaccharide (LPS), and white spot syndrome virus (WSSV), thus, suggesting its probable involvement in shrimp immune response. Moreover, knockdown of LvPim1 resulted in increased hemocytes apoptosis; shown by high caspase3/7 activity, coupled with increase in pro-apoptotic LvCaspase3 and LvCytochrome C, and decrease in pro-survival LvBcl2, LvIAP1, and LvIAP2 mRNA expression in hemocytes. Finally, LvPim1 knockdown renders shrimps more susceptible to V. parahaemolyticus infection. Taken together, our present data strongly suggest that LvPim1 is involved in modulating shrimp resistance to pathogen infection, promote hemocytes survival, and therefore plays a role in shrimp immune response.
丝氨酸/苏氨酸激酶 Pim1 与多种细胞功能相关，包括脊椎动物中的增殖、存活、分化、凋亡、肿瘤发生、免疫调节和炎症。然而，关于 Pim1 在无脊椎动物免疫中的作用知之甚少。在这项研究中，我们首次在凡纳滨对虾中鉴定并表征了一个 Pim1（LvPim1）基因，其全长 cDNA 为 2352bp，1119bp 的开放阅读框（ORF）编码一个推定的 372 个氨基酸的蛋白质，其中包含一个典型的丝氨酸/苏氨酸激酶结构域。序列和系统发育分析表明，LvPim1 与脊椎动物的 Pim1 具有密切的进化关系。实时 qPCR 分析显示，LvPim1 在所有检测的组织中广泛表达；其在肝胰腺和血细胞中受到副溶血弧菌、链球菌、脂多糖（LPS）和白斑综合征病毒（WSSV）的刺激后，其转录水平被诱导，因此，表明其可能参与虾的免疫反应。此外，LvPim1 的敲低导致血细胞凋亡增加；通过高半胱天冬酶 3/7 活性，加上促凋亡的 LvCaspase3 和 LvCytochrome C 的增加，以及抗凋亡的 LvBcl2、LvIAP1 和 LvIAP2mRNA 表达的减少来证明。最后，LvPim1 的敲低使虾更容易受到副溶血弧菌的感染。总之，我们目前的数据强烈表明，LvPim1 参与调节虾对病原体感染的抵抗力，促进血细胞存活，因此在虾的免疫反应中发挥作用。
