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表达谱和相互作用表明,在大黄鱼(Larimichthys crocea)中,TBK1可受Nrdp1调控以响应免疫刺激。

Expression profiles and interaction suggest TBK1 can be regulated by Nrdp1 in response to immune stimulation in large yellow croaker Larimichthys crocea.

作者信息

Zhang Dong Ling, Yu Da Hui, Chen Jian, Fan Sigang, Wang Zhi Yong

机构信息

Key Laboratory of Healthy Mariculture for The East China Sea, Ministry of Agriculture, Fisheries College, Jimei University, Xiamen 361021, PR China.

South China Sea Resource Exploitation and Protection Collaborative Innovation Center (SCS-REPIC), South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou, 510300, PR China.

出版信息

Fish Shellfish Immunol. 2015 Oct;46(2):745-52. doi: 10.1016/j.fsi.2015.08.013. Epub 2015 Aug 17.

Abstract

TBK1 has been extensively studied in mammals because of its important roles as a molecular bridge, linking the TLRs (TLR3 and TLR4) and RLRs signals to activate transcriptional factors IRF3 and IRF7 for IFN-I production. However, the information on molecular and functional characteristics of TBK1 in teleosts is limited. In this study, the molecular characterization and immune response of TBK1 in Larimichthys crocea (named as LcTBK1) as well as its interaction with Nrdp1 were investigated. Sequence analysis demonstrated that LcTBK1 included four functional motifs, the N-terminal protein kinase domain and ATP-binding site, middle ULD and C-terminal coiled-coil domain. The tissue expression profiles indicated that LcTBK1 gene was constitutively expressed in the twelve tissues examined, with high expression in brain. Temporal expression analysis showed that LcTBK1 mRNA was obviously increased in the liver after injection of LPS, Poly I:C and inactive Vibrio parahaemolyticus, however, declined at some time points in spleen and head-kidney. Furthermore, we found that LcTBK1 can interact with LcNrdp1, an E3 ubiquitin ligase that involved in immune response to Cryptocaryon irritans infection in L. crocea. The qPCR showed that LcNrdp1 was also significantly up-regulated in liver, down-regualted at some time points in spleen and head-kidney after LPS, Poly I:C and inactive V. parahaemolyticus injection, although the expression patterns of the two genes after the three treatments were different in change magnitude and up-regulation timespan. These results suggested that LcTBK1 was involved in L. crocea defense against the pathogen infection and can be regulated by Nrdp1 in PPRs signaling pathway of fishes.

摘要

由于TBK1作为分子桥梁发挥着重要作用,将Toll样受体(TLR3和TLR4)和视黄酸诱导基因样受体(RLR)信号联系起来,激活转录因子IRF3和IRF7以产生I型干扰素,因此在哺乳动物中已对其进行了广泛研究。然而,关于硬骨鱼中TBK1的分子和功能特性的信息有限。在本研究中,对大黄鱼中TBK1(命名为LcTBK1)的分子特征、免疫反应及其与Nrdp1的相互作用进行了研究。序列分析表明,LcTBK1包含四个功能基序,即N端蛋白激酶结构域和ATP结合位点、中间的ULD以及C端卷曲螺旋结构域。组织表达谱表明,LcTBK1基因在所检测的12个组织中组成性表达,在脑中表达量高。时序表达分析表明,注射脂多糖(LPS)、聚肌胞苷酸(Poly I:C)和灭活的副溶血性弧菌后,LcTBK1 mRNA在肝脏中明显增加,但在脾脏和头肾的某些时间点下降。此外,我们发现LcTBK1可以与LcNrdp1相互作用,LcNrdp1是一种E3泛素连接酶,参与大黄鱼对刺激隐核虫感染的免疫反应。定量PCR显示,注射LPS、Poly I:C和灭活的副溶血性弧菌后,LcNrdp1在肝脏中也显著上调,在脾脏和头肾的某些时间点下调,尽管这两个基因在三种处理后的表达模式在变化幅度和上调时间跨度上有所不同。这些结果表明,LcTBK1参与大黄鱼对病原体感染的防御,并且在鱼类的模式识别受体(PPR)信号通路中可受Nrdp1调控。

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