Molecular Cloning and Effects of Tm14-3-3ζ-Silencing on Larval Survivability Against E. coli and C. albicans in Tenebrio molitor.
Molecular Cloning and Effects of Tm14-3-3ζ-Silencing on Larval Survivability Against E. coli and C. albicans in Tenebrio molitor.
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TM14-3-3ζ分解对Tenebrio molitor中针对大肠杆菌和白色念珠菌的幼虫生存能力的分子克隆和影响。
DOI：
10.3390/genes9070330
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发表时间：
2018-06-29
期刊：
影响因子：
3.5
通讯作者：
Han YS
中科院分区：
文献类型：
--
作者：
Seong JH;Jo YH;Seo GW;Park S;Park KB;Cho JH;Ko HJ;Kim CE;Patnaik BB;Jun SA;Choi YS;Kim YW;Bang IS;Lee YS;Han YS
14-3-3蛋白家族在磷酸化依赖的信号通路中发挥关键的调节作用，包括细胞存活和增殖、细胞凋亡、染色质结构调节和自噬。本研究从鞘翅目昆虫黄颡鱼（Tenebrio molitor）的表达序列标签（ESTs）和RNA测序（RNA- seq）数据库中鉴定了14-3-3蛋白的zeta亚型（命名为Tm14-3-3ζ）。Tm14-3-3ζ信使RNA （mRNA）在幼虫期和成虫期的免疫器官中表达水平较高，并在感染大肠杆菌和白色念珠菌后3小时内表现出近5倍的诱导。为了研究Tm14-3-3ζ的生物学功能，制备了一种基于多肽的Tm14-3-3ζ多克隆抗体，并用Western blot分析证实了该抗体的特异性。免疫染色和共聚焦显微镜分析表明，Tm14-3-3ζ主要表达于中肠上皮细胞的细胞膜、脂肪体的细胞核和血细胞的胞浆中。基因沉默Tm14-3-3ζ增加了感染大肠杆菌和白色念珠菌后7天的幼虫死亡率。我们的研究结果表明，T. molitor中的14-3-3ζ在宿主对细菌和真菌的防御机制中是必不可少的。
The 14-3-3 family of proteins performs key regulatory functions in phosphorylation-dependent signaling pathways including cell survival and proliferation, apoptosis, regulation of chromatin structure and autophagy. In this study, the zeta isoform of 14-3-3 proteins (designated as Tm14-3-3ζ) was identified from the expressed sequence tags (ESTs) and RNA sequencing (RNA-Seq) database of the coleopteran pest, Tenebrio molitor. Tm14-3-3ζ messenger RNA (mRNA) is expressed at higher levels in the immune organs of the larval and adult stages of the insect and exhibit almost five-fold induction within 3 h post-infection of the larvae with Escherichia coli and Candida albicans. To investigate the biological function of Tm14-3-3ζ, a peptide-based Tm14-3-3ζ polyclonal antibody was generated in rabbit and the specificity was confirmed using Western blot analysis. Immunostaining and confocal microscopic analyses indicate that Tm14-3-3ζ is mainly expressed in the membranes of midgut epithelial cells, the nuclei of fat body and the cytosol of hemocytes. Gene silencing of Tm14-3-3ζ increases mortality of the larvae at 7 days post-infection with E. coli and C. albicans. Our findings demonstrate that 14-3-3ζ in T. molitor is essential in the host defense mechanisms against bacteria and fungi.
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