RAT GENE ENCODING THE 78-KDA GLUCOSE-REGULATED PROTEIN GRP78 - ITS REGULATORY SEQUENCES AND THE EFFECT OF PROTEIN GLYCOSYLATION ON ITS EXPRESSION
RAT GENE ENCODING THE 78-KDA GLUCOSE-REGULATED PROTEIN GRP78 - ITS REGULATORY SEQUENCES AND THE EFFECT OF PROTEIN GLYCOSYLATION ON ITS EXPRESSION
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DOI：
10.1073/pnas.84.3.680
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发表时间：
1987-02-01
影响因子：
11.1
通讯作者：
LEE, AS
中科院分区：
文献类型：
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作者：
CHANG, SC;WOODEN, SK;LEE, AS
78-kDa葡萄糖调节蛋白GRP 78是一种在动物细胞中广泛表达的应激诱导蛋白。在本文中，我们表明，这种内质网定位的蛋白质的第一个外显子由一个18个氨基酸的前导序列，富含疏水残基，其次是一个高度酸性的成熟的N-末端和11个氨基酸的结构域，是由70 kDa的热休克蛋白家族的成员共享。这个共有结构域的末端也标志着该基因第一个内含子的开始。通过缺失分析确定了对钙离子载体诱导和温度敏感性突变重要的启动子元件上游的DNA区域。我们的研究结果表明，从85到480个核苷酸的主要转录起始位点上游跨越区域是重要的两个诱导条件。有证据表明，蛋白质糖基化的扰动可能是参与GRP转录激活的常见刺激之一，我们测量了A23187、葡萄糖饥饿和温度变化诱导条件下的糖基化速率。糖基化速率和GRP 78转录物的稳态水平之间观察到的负相关支持了这一假设。
The 78-kDa glucose-regulated protein GRP78 is a stress-inducible protein ubiquitously expressed in animal cells. In this paper we show that the first exon of this endoplasmic reticulum-localized protein consists of an 18 amino acid leader sequence rich in hydrophobic residues, followed by a highly acidic mature N-terminus and an 11 amino acid domain that is shared by members of the 70-kDa heat shock protein family. The end of this shared domain also marks the beginning of the first intron of this gene. A DNA region upstream of the promoter element important for induction by calcium ionophore and by a temperature-sensitive mutation was identified by deletion analysis. Our results indicate that a region spanning from 85 to 480 nucleotides upstream of the major transcription initiation site is important for both induction conditions. With evidence suggesting that perturbations in protein glycosylation may be one of the common stimuli involved in transcription activation of the GRPs, we measured the rate of glycosylation during A23187, glucose starvation, and temperature-shift induced conditions. The inverse correlation observed between the rate of glycosylation and the steady-state level of the GRP78 transcripts lends support to this hypothesis.