Isolation and analysis of the expression of two genes for the 81-kilodalton heat-shock proteins from Arabidopsis.
Isolation and analysis of the expression of two genes for the 81-kilodalton heat-shock proteins from Arabidopsis.
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DOI：
10.1104/pp.99.2.383
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发表时间：
1992-06
期刊：
影响因子：
7.4
通讯作者：
Taku Takahashi;Satoshi Naito;Y. Komeda
中科院分区：
文献类型：
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作者：
Taku Takahashi;Satoshi Naito;Y. Komeda
我们已经克隆和表征了两个成员的基因家族的81千道尔顿热休克蛋白从拟南芥，HSP 81 -1和HSP 81 -2。将HSP 81 -1基因的整个基因组序列与先前报道为AtHS 83的相应全长cDNA（TW Conner，PR LaFayette，RT Nagao，JL Key [1990] Plant Physiol 94：1689-1695）进行比较，揭示存在315、83和88个碱基对的三个内含子。与此相反，HSP 81 -2基因组和部分cDNA序列的分析表明，HSP 81 -2基因被中断只有两个内含子的304和106碱基对。根据S1核酸酶的实验结果绘制了两种相应mRNA的5 ′-起始位点。这两个基因编码的蛋白质的推导的氨基酸序列显示出88%的同一性，并且它们也显示出与酵母和动物细胞中的hsp 90蛋白质家族惊人的相似性。从转录物的北方印迹分析结果来看，在没有热休克的情况下，HSP 81 -1基因的表达似乎仅以非常低的水平发生，并且受到热（35摄氏度）的强烈诱导。HSP 81 -2基因以高得多的水平组成型表达，并且其表达通过升高的温度适度增强。严重的热休克似乎阻断了从HSP 81 -1转录的前体mRNA的剪接。我们还研究了砷和镉对HSP 81基因表达的影响，以及对拟南芥中其他组的热休克蛋白基因。镉处理是轻微有效的诱导HSP基因，而砷胁迫强烈刺激每个mRNA的积累在一个协调的方式。
We have cloned and characterized two members of the family of genes for the 81-kilodalton heat-shock proteins from Arabidopsis thaliana, HSP81-1 and HSP81-2. Comparison of the entire genomic sequence of the HSP81-1 gene with the corresponding full-length cDNA previously reported as AtHS83 (TW Conner, PR LaFayette, RT Nagao, JL Key [1990] Plant Physiol 94: 1689-1695) reveals the presence of three introns of 315, 83, and 88 base pairs. By contrast, analysis of the HSP81-2 genomic and partial cDNA sequences suggests that the HSP81-2 gene is interrupted by only two introns of 304 and 106 base pairs. The 5'-initiation sites of the two corresponding mRNAs were mapped from results of experiments with S1 nuclease. The deduced amino acid sequences of the proteins encoded by these two genes show 88% identity, and they also show striking similarities to the hsp90 family of proteins in yeast and animal cells. From the results of northern blot analysis of transcripts, it appears that the expression of the HSP81-1 gene occurs at only very low levels in the absence of heat shock and is strongly induced by heat (35 degrees C). The HSP81-2 gene is constitutively expressed at much higher levels, and its expression is moderately enhanced by elevated temperatures. Severe heat shock appears to block the splicing of the pre-mRNA transcribed from HSP81-1. We also examined the effects of arsenite and cadmium on the expression of the HSP81 genes, as well as on other groups of hsp genes in Arabidopsis. Treatment with cadmium was marginally effective in inducing hsp genes, whereas arsenite stress strongly stimulated the accumulation of each mRNA in a coordinated fashion.