Song Yu-Feng, Luo Zhi, Huang Chao, Chen Qi-Liang, Pan Ya-Xiong, Xu Yi-Huan
Key Laboratory of Freshwater Animal Breeding, Ministry of Agriculture, Fishery College, Huazhong Agricultural University, Wuhan 430070, China Freshwater Aquaculture Collaborative Innovative Center of Hubei Province, Wuhan 430070, China.
Key Laboratory of Freshwater Animal Breeding, Ministry of Agriculture, Fishery College, Huazhong Agricultural University, Wuhan 430070, China Freshwater Aquaculture Collaborative Innovative Center of Hubei Province, Wuhan 430070, China
G3 (Bethesda). 2015 Aug 13;5(10):2091-104. doi: 10.1534/g3.115.019950.
Two endoplasmic reticulum (ER) molecular chaperones [glucose-regulated protein 78 (grp78) and calreticulin (crt)] and three ER stress sensors [PKR-like ER kinase (perk), inositol requiring enzyme (ire)-1α, and activating transcription factor (atf)-6α] cDNAs were first characterized from yellow catfish, Pelteobagrus fulvidraco. The predicted amino acid sequences for the yellow catfish grp78, crt, perk, ire-1α, and atf-6α revealed that the proteins contained all of the structural features that were characteristic of the five genes in other species, including the KDEL motif, signal peptide, sensor domain, and effector domain. mRNAs of the five genes mentioned above were expressed in various tissues, but their mRNA levels varied among tissues. Dietary Cu excess, but not Cu deficiency, activated the chaperones (grp78 and crt) and folding sensors in ER, and the UPR signaling pathways (i.e., perk-eif2α and the ire1-xbp1) in a tissue-specific manner. For the first time, our study cloned grp78, crt, perk, ire-1α, and atf-6α genes in yellow catfish and demonstrated their differential expression among tissues. Moreover, the present study also indicated differential regulation of these ER stress-related genes by dietary Cu deficiency and excess, which will be beneficial for us to evaluate effects of dietary Cu levels in fish at the molecular level, based on the upstream pathway of lipid metabolism (the ER) and thus provide novel insights regarding the nutrition of Cu in fish.
首次从黄颡鱼(Pelteobagrus fulvidraco)中鉴定出两种内质网(ER)分子伴侣[葡萄糖调节蛋白78(grp78)和钙网蛋白(crt)]以及三种ER应激传感器[蛋白激酶R样内质网激酶(perk)、肌醇需求酶(ire)-1α和激活转录因子(atf)-6α]的cDNA。黄颡鱼grp78、crt、perk、ire-1α和atf-6α的预测氨基酸序列显示,这些蛋白质包含了其他物种中五个基因所特有的所有结构特征,包括KDEL基序、信号肽、传感器结构域和效应器结构域。上述五个基因的mRNA在各种组织中均有表达,但其mRNA水平在不同组织间存在差异。日粮铜过量而非铜缺乏会以组织特异性方式激活内质网中的分子伴侣(grp78和crt)和折叠传感器以及未折叠蛋白反应信号通路(即perk-eif2α和ire1-xbp1)。本研究首次克隆了黄颡鱼的grp78、crt、perk、ire-1α和atf-6α基因,并证明了它们在不同组织中的差异表达。此外,本研究还表明日粮铜缺乏和过量对这些内质网应激相关基因的调控存在差异,这将有助于我们基于脂质代谢的上游途径(内质网)在分子水平上评估日粮铜水平对鱼类的影响,从而为鱼类铜营养提供新的见解。