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古菌类TATA样启动子的热激诱导性受相邻序列元件控制。

Heat shock inducibility of an archaeal TATA-like promoter is controlled by adjacent sequence elements.

作者信息

Thompson D K, Daniels C J

机构信息

Department of Microbiology, The Ohio State University, Columbus 43210, USA.

出版信息

Mol Microbiol. 1998 Feb;27(3):541-51. doi: 10.1046/j.1365-2958.1998.00698.x.

Abstract

The expression of a heat-inducible cct1 (chaperonin-containing Tcp-1) family member gene is regulated at the transcription level in the archaeon Haloferax volcanii. Transcriptional fusions of the cct1 promoter region with a yeast proline tRNA reporter gene were constructed to analyse the functional domains of this archaeal heat shock promoter. Both basal and heat-induced transcription of the reporter gene was directed by an archaeal consensus TATA element (5'-TTTATA-3') centred 25bp upstream of the transcription start site. Deletion mutagenesis indicated that the 5' boundary of the cct1 regulatory region mapped to position -37. Nucleotide alignment with the 5' flanking regions of two additional cct-related genes identified in H. volcanii showed a high degree of sequence conservation between positions +1 and -37, especially in and immediately surrounding the TATA element of the putative core promoter. Mutational analysis of conserved sequences demonstrated that basal and heat-induced transcription required sequence elements located upstream and downstream of the TATA-box. These findings indicate that the regulatory sequences involved in heat-induced transcription lie within the core promoter region and suggest that the mechanism controlling heat shock gene expression in H. volcanii differs from the bacterial and eukaryal strategies.

摘要

在古菌嗜盐嗜热栖热菌中,一个热诱导型cct1(含伴侣蛋白的Tcp-1)家族成员基因的表达在转录水平受到调控。构建了cct1启动子区域与酵母脯氨酸tRNA报告基因的转录融合体,以分析该古菌热休克启动子的功能结构域。报告基因的基础转录和热诱导转录均由位于转录起始位点上游25bp处的一个古菌共有TATA元件(5'-TTTATA-3')指导。缺失诱变表明,cct1调控区域的5'边界定位于-37位。与在嗜盐嗜热栖热菌中鉴定出的另外两个cct相关基因的上游侧翼区域进行核苷酸比对,结果显示在+1和-37位之间具有高度的序列保守性,尤其是在假定核心启动子的TATA元件内部及紧邻区域。对保守序列的突变分析表明,基础转录和热诱导转录需要位于TATA框上游和下游的序列元件。这些发现表明,参与热诱导转录的调控序列位于核心启动子区域内,并且提示嗜盐嗜热栖热菌中控制热休克基因表达的机制不同于细菌和真核生物的策略。

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