Human HSP70 promoter contains at least two distinct regulatory domains.-猫眼课题宝

人类 HSP70 启动子包含至少两个不同的调控域。

DOI：

10.1073/pnas.83.3.629

发表时间：

1986

期刊：

Proceedings of the National Academy of Sciences of the United States of America

影响因子：

11.1

通讯作者：

中科院分区：

综合性期刊1区

文献类型：

作者：

Barbara J. Wu;R. Kingston;R. Morimoto

人类HSP70基因的表达受多种生理应激的诱导，包括热休克和重金属的暴露，或在非应激条件下，如对血清刺激的反应。我们以前已经证明，在任何一种情况下，受调控的表达都处于转录的初级水平。为了确定转录是通过单个还是多个遗传元件介导的，我们通过构建含有不同数量的5‘侧翼区的嵌合基因来解剖人类HSP70基因转录起始点上游的序列，这些嵌合基因与编码氯霉素乙酰转移酶的细菌基因融合。通过对单独稳定的转染体进行S1核酸酶分析，确定嵌合基因的转录。热休克和镉诱导所需的序列介于-107和-68之间。在这个区域内有CTGGAATATTCCCG序列，它在12/14位与果蝇热休克基因的热休克元件相同，以及一个单独的序列CGNCCCGG，它与人类金属硫蛋白II金属反应元件的核心同源。血清刺激转录所需的序列不同于热休克元件。CCAAT at-68序列是高水平正确启动的转录本所必需的，而富含嘌呤的序列GAAGGGAAAAG，at-58是血清刺激所必需的。人类HSP70启动子至少包含两个调控结构域--对热休克或镉反应的远端结构域和对血清刺激反应的近端结构域。

The expression of the human HSP70 gene is induced by a wide range of physiological stresses, including exposure to heat shock and heavy metals, or under nonstress conditions, such as in response to serum stimulation. We have previously demonstrated that in either case the regulated expression is at the primary level of transcription. To determine whether transcription is mediated through a single or multiple genetic elements, we have dissected the sequences upstream of the transcription start site of the human HSP70 gene by constructing chimeric genes retaining variable amounts of 5' flanking regions fused to the bacterial gene encoding chloramphenicol acetyltransferase. Transcription from the chimeric genes was determined by S1 nuclease analysis of separate stable transfectants. The sequences required for heat shock and cadmium induction lie between -107 and -68. Within this region is the sequence CTGGAATATTCCCG, which is identical in 12/14 positions with the heat shock element of Drosophila heat shock genes, and a separate sequence, CGNCCCGG, which is homologous to the core of the human metallothionein II metal-responsive element. The sequences required for serum-stimulated transcription are distinct from the heat shock element. The sequence CCAAT at -68 is required for high levels of correctly initiated transcripts, and a purine-rich sequence, GAAGGGAAAAG, at -58 is required for serum stimulation. The human HSP70 promoter contains at least two regulatory domains--a distal domain responsive to heat shock or cadmium and a proximal domain responsive to stimulation by serum.

Adenovirus EIIA early promoter: transcriptional control elements and induction by the viral pre-early EIA gene, which appears to be sequence independent.