Isolation of a Latimeria menadoensis heat shock protein 70 (Lmhsp70) that has all the features of an inducible gene and encodes a functional molecular chaperone.
Isolation of a Latimeria menadoensis heat shock protein 70 (Lmhsp70) that has all the features of an inducible gene and encodes a functional molecular chaperone.
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分离出 Latimeria menadoensis 热休克蛋白 70 (Lmhsp70)，该蛋白具有诱导基因的所有特征，并编码功能性分子伴侣。
DOI：
10.1007/s00438-009-0456-4
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发表时间：
2009
期刊：
影响因子：
--
通讯作者：
Blatch,GregoryL
中科院分区：
文献类型：
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作者：
Modisakeng,KeoagileW;Jiwaji,Meesbah;Pesce,Eva-Rachele;Robert,Jacques;Amemiya,ChrisT;Dorrington,RosemaryA;Blatch,GregoryL
分子伴侣有助于其他蛋白质的正确折叠，热休克蛋白是分子伴侣的主要类型之一。热休克蛋白70(Hsp70)已被广泛研究，并被证明在几乎所有已研究的原核和真核系统中对细胞蛋白质的动态平衡起着至关重要的作用。由于对腔棘鱼伴侣蛋白的研究非常有限，本研究的主要目的是对腔棘鱼Hsp70的遗传学和生化特征进行研究。我们成功地克隆了腔棘鱼hsp70基因Lmhsp70，发现它含有一个无内含子的编码区和一个潜在的上游调控区。Lmhsp70根据保守的结构和功能编码了一个典型的hsp70，预测的上游调控区包含六个潜在的启动子元件和三个潜在的热休克元件(HSE)。编码区的无内含子性质和HSE的存在表明Lmhsp70是应激诱导的。系统发育分析提供了进一步的证据，证明Lmhsp70可能是可诱导的，并且作为硬骨鱼和四足动物之间的分支中间体。重组LmHsp70被成功地超量生产和纯化，并通过ATPase活性测定证明其具有功能。综上所述，这些数据首次提供了腔棘鱼编码功能分子伴侣系统的证据。
Molecular chaperones facilitate the correct folding of other proteins, and heat shock proteins form one of the major classes of molecular chaperones. Heat shock protein 70 (Hsp70) has been extensively studied, and shown to be critically important for cellular protein homeostasis in almost all prokaryotic and eukaryotic systems studied to date. Since there have been very limited studies conducted on coelacanth chaperones, the main objective of this study was to genetically and biochemically characterize a coelacanth Hsp70. We have successfully isolated an Indonesian coelacanth (L. menadoensis)hsp70gene,Lmhsp70, and found that it contained an intronless coding region and a potential upstream regulatory region.Lmhsp70encoded a typical Hsp70 based on conserved structural and functional features, and the predicted upstream regulatory region was found to contain six potential promoter elements, and three potential heat shock elements (HSEs). The intronless nature of the coding region and the presence of HSEs suggested thatLmhsp70was stress-inducible. Phylogenetic analyses provided further evidence thatLmhsp70was probably inducible, and that it branched as a clade intermediate between bony fish and tetrapods. Recombinant LmHsp70 was successfully overproduced, purified and found to be functional using ATPase activity assays. Taken together, these data provide evidence for the first time that the coelacanth encodes a functional molecular chaperone system.