Characterization of multiple members of the HSP70 family in platyfish culture cells: molecular evolution of stress protein HSP70 in vertebrates.
Characterization of multiple members of the HSP70 family in platyfish culture cells: molecular evolution of stress protein HSP70 in vertebrates.
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DOI：
10.1016/j.gene.2004.04.023
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发表时间：
2004-07
期刊：
影响因子：
3.5
通讯作者：
M. Yamashita;K. Hirayoshi;K. Nagata
中科院分区：
文献类型：
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作者：
M. Yamashita;K. Hirayoshi;K. Nagata
将培养的对虾成纤维细胞系EHS细胞(对虾成纤维细胞系)的培养温度从28℃转变到37℃，诱导了一系列应激蛋白。双向凝胶电泳(2D-PAGE)分析表明，细胞表达3种不同的热休克蛋白70(HSP70)家族蛋白：热诱导型HSP70，结构性表达的热休克同源蛋白70(HSC70)及其磷酸化异构体，以及葡萄糖调节蛋白78(GRP78)。从热诱导型HSP70的两个主要亚型HSP70-1和HSP70-2以及HSC70的基因文库中分离到3个不同的克隆。我们比较了贝类HSP70和HSC70蛋白与其他脊椎动物HSP70和HSC70蛋白推导的氨基酸序列。系统进化分析表明，脊椎动物HSP70可分为四类：(A)鱼类HSP70，鱼类HSP70-1和HSP70-2两种亚型；(B)哺乳动物睾丸特异的HSP70相关蛋白；(C)哺乳动物热诱导HSP70B‘；以及(D)哺乳动物主要组织相容性复合体(MHC)连锁的HSP70，包括MHC连锁的热诱导HSP70和睾丸特异的HSP70相关蛋白。这些发现表明，脊椎动物HSP70是在脊椎动物进化过程中起源于单一的祖先HSP70基因，而在鱼类和高等脊椎动物的遗传分化过程中，可能进化了多个副本的热诱导HSP70。
A shift from 28 to 37 °C in the incubation temperature of a culture of the platyfish fibroblast cell line, EHS cells (platyfish fibroblast cell line), induced a set of stress proteins. A two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) analysis showed that the cells expressed three genetically distinct forms of heat-shock protein 70 (HSP70) family proteins: heat-inducible forms of HSP70, the constitutively expressed heat-shock cognate protein 70 (HSC70) and its phosphorylated isoform, and the glucose-regulated protein 78 (GRP78). Three different clones encoding two major isoforms of heat-inducible HSP70, platyfish HSP70-1 and HSP70-2, and of the HSC70 were isolated from a platyfish cDNA library. We compared the deduced amino acid sequences of the platyfish HSP70 and HSC70 proteins with those of other vertebrates. Phylogenetic analysis showed that vertebrate HSP70 could be classified into four cluster groups: (a) fish HSP70, with two isoforms of heat-inducible HSP70 in fish, fish HSP70-1 and HSP70-2; (b) the mammalian testis-specific HSP70-related protein HST70; (c) the mammalian heat-inducible HSP70B'; and (d) the mammalian major histocompatibility complex (MHC)-linked HSP70, including the MHC-linked heat-inducible HSP70 and the testis-specific HSP70-related protein. These findings suggest that vertebrate HSP70 was derived from a single ancestral HSP70 gene during vertebrate evolution and that multiple copies of heat-inducible HSP70 were probably evolved during genetic divergence in fish and higher vertebrates.