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植物转录因子TGA1可刺激酿酒酵母中CaMV 35S启动子的表达。

The plant transcription factor TGA1 stimulates expression of the CaMV 35S promoter in Saccharomyces cerevisiae.

作者信息

Rüth J, Schweyen R J, Hirt H

机构信息

Institute of Microbiology and Genetics, University of Vienna, Austria.

出版信息

Plant Mol Biol. 1994 May;25(2):323-8. doi: 10.1007/BF00023248.

Abstract

We have previously shown that two CRE elements situated on a 31 bp region of the cauliflower mosaic virus (CaMV) 35S promoter activate gene expression in the yeast Saccharomyces cerevisiae and are regulated by cAMP. Studies with the yeast transcription factors GCN4, SKO1 and YAP1, which bind CRE-like sequences, showed no influence on expression of the 35S promoter indicating that a yet unknown factor is involved in activation. Band shift experiments with the 31 bp promoter region revealed binding of similar factors in yeast and plant protein extracts. In a previous study this promoter region was shown to confer tissue-specific expression in plants and to interact with the transcription factor TGA1. To test whether expression of TGA1 in yeast also stimulates transcription of the 35S promoter, we co-transformed yeast cells with a cDNA clone of this transcription factor and a 35S promoter/reporter gene construct. Promoter activity studies revealed that TGA1 confers enhanced expression of a reporter gene under the control of the 35S promoter in yeast cells. Yeast cells that were transformed with a 35S promoter construct that containing a mutated TGA1-binding site showed that both TGA1 and the intact binding site are necessary for this activation. These results suggest that stimulation of the 35S promoter by TGA1 is mediated by competition with an endogenous down-regulating yeast factor that is modulated by the nutritional state of the cells.

摘要

我们之前已经表明,位于花椰菜花叶病毒(CaMV)35S启动子31 bp区域的两个CRE元件可激活酿酒酵母中的基因表达,并受cAMP调控。对结合CRE样序列的酵母转录因子GCN4、SKO1和YAP1的研究表明,它们对35S启动子的表达没有影响,这表明有一个未知因子参与激活过程。对31 bp启动子区域进行的凝胶迁移实验显示,酵母和植物蛋白提取物中存在类似因子的结合。在之前的一项研究中,该启动子区域被证明可赋予植物组织特异性表达,并与转录因子TGA1相互作用。为了测试TGA1在酵母中的表达是否也能刺激35S启动子的转录,我们将该转录因子的cDNA克隆与35S启动子/报告基因构建体共转化酵母细胞。启动子活性研究表明,TGA1可增强酵母细胞中35S启动子控制下报告基因的表达。用含有突变TGA1结合位点的35S启动子构建体转化的酵母细胞表明,TGA1和完整的结合位点对于这种激活都是必需的。这些结果表明,TGA1对35S启动子的刺激是通过与一种受细胞营养状态调节的内源性酵母下调因子竞争来介导的。

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