College of Life Science, Nanchang University, Nanchang, 330031, Jiangxi, China.
State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, 330047, Jiangxi, China.
Dev Comp Immunol. 2021 Mar;116:103909. doi: 10.1016/j.dci.2020.103909. Epub 2020 Oct 29.
Mre11A is considered as a cytosolic DNA receptor in mammals. However, it is rarely known about Mre11A in other vertebrates. Recently, a mammalian ortholog of Mre11A has been identified in grass carp (Ctenopharyngodon idellus) in our lab. Phylogenetic-tree analysis provided evidence for a close genetic relationship between C.idellus Mre11A and Carassius auratus Mre11A. The tissue expression profile of CiMre11A was detected, with a relatively higher level of expression in kidney, intestines, liver and spleen than that in other tissues after grass carp reovirus (GCRV) infection. Similarly, CiMre11A was also up-regulated in CIK cells after treatment with GCRV. Q-PCR and dual-luciferase assays indicated that the transcription levels of IFN1 and ISG15 were inhibited by CiMre11A knockdown, but were gradually augmented after CIK cells were transfected with increasing amounts of CiMre11A. Subcellular localization assays showed that a part of CiMre11A was translocated from the nucleus to the cytoplasm. Co-immunoprecipitation and co-localization assays demonstrated that CiMre11A interacts with CiSTING in response to GCRV infection. In CIK cells, the expressions of both IFN1 and ISG15 were acutely up-regulated by CiMre11A overexpression, as well as by co-overexpression of CiMre11A and CiSTING. CiMre11A and CiSTING induced the phosphorylation and cytoplasmic-to-nuclear translocation of IRF7 in CIK cells. The multiplication of GCRV in CIK cells was inhibited by the overexpression of CiMre11A and CiSTING.
Mre11A 被认为是哺乳动物细胞质中的 DNA 受体。然而,关于其他脊椎动物中的 Mre11A 却知之甚少。最近,我们实验室在草鱼(Ctenopharyngodon idellus)中鉴定出了一种 Mre11A 的哺乳动物同源物。系统进化树分析为草鱼 Mre11A 和鲤鱼 Mre11A 之间的密切遗传关系提供了证据。检测了 CiMre11A 的组织表达谱,发现草鱼呼肠孤病毒(GCRV)感染后,肾脏、肠道、肝脏和脾脏中的表达水平相对较高。同样,CIK 细胞经 GCRV 处理后,CiMre11A 也被上调。Q-PCR 和双荧光素酶报告基因检测表明,CiMre11A 敲低抑制了 IFN1 和 ISG15 的转录水平,但随着 CIK 细胞转染 CiMre11A 的量逐渐增加,转录水平逐渐升高。亚细胞定位实验表明,一部分 CiMre11A 从细胞核转移到细胞质。免疫共沉淀和共定位实验表明,CiMre11A 与 CiSTING 相互作用,响应 GCRV 感染。在 CIK 细胞中,CiMre11A 过表达以及与 CiSTING 共表达均可急性上调 IFN1 和 ISG15 的表达。CiMre11A 和 CiSTING 诱导 CIK 细胞中 IRF7 的磷酸化和从细胞质到细胞核的转位。CIK 细胞中 GCRV 的增殖被 CiMre11A 和 CiSTING 的过表达所抑制。