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酵母甘油醛-3-磷酸脱氢酶(GAPDH)启动子的上游激活序列(UAS)由多个一般功能元件组成,包括RAP1和GRF2结合位点。

The UAS of the yeast GAPDH promoter consists of multiple general functional elements including RAP1 and GRF2 binding sites.

作者信息

Yagi S, Yagi K, Fukuoka J, Suzuki M

机构信息

Corporate Research & Development Laboratory, Saitama, Japan.

出版信息

J Vet Med Sci. 1994 Apr;56(2):235-44. doi: 10.1292/jvms.56.235.

Abstract

The upstream activating sequence (UAS) of TDH3, one of three genes encoding glyceraldehyde phosphate dehydrogenase in Saccharomyces cerevisiae, was characterized by using a series of external and internal deletion mutants of the TDH3 upstream region. The levels of activation by these deletions of transcription mediated through either the segment of TDH3 promoter or the segment of ADH1 (alcohol dehydrogenase 1 gene) promoter were quantitatively examined and the region between -583 and -447 was found to be required for full transcriptional activation with either promoter segment. It has been demonstrated that the protein binding site involved in the formation of two DNA-protein complexes is identical with the consensus RAP1 binding sequence by methylation interference assay. Surprisingly, the UAS fragment composed of the 22-mer sequence containing exclusively a RAP1 binding sequence showed full activation, suggesting that the RAP1-dependent transcriptional activation is a primary positive control in the TDH3 gene expression. In addition, a pair of inverted repeat sequences homologous to the binding sequence for GRF2, another yeast trans-acting factor, and directly repeated sequences containing a CATCC motif were also found upstream and downstream, respectively, of the RAP1 binding site. Deletion analysis suggested that these elements could also function as regulatory elements for transcription.

摘要

TDH3是酿酒酵母中编码磷酸甘油醛脱氢酶的三个基因之一,其上游激活序列(UAS)通过使用TDH3上游区域的一系列外部和内部缺失突变体进行了表征。对这些通过TDH3启动子片段或ADH1(乙醇脱氢酶1基因)启动子片段介导的转录缺失的激活水平进行了定量检测,发现-583至-447区域是使用任一启动子片段进行完全转录激活所必需的。通过甲基化干扰试验已证明,参与形成两种DNA-蛋白质复合物的蛋白质结合位点与共有RAP1结合序列相同。令人惊讶的是,由仅包含一个RAP1结合序列的22聚体序列组成的UAS片段显示出完全激活,这表明依赖RAP1的转录激活是TDH3基因表达中的主要正调控。此外,在RAP1结合位点的上游和下游分别还发现了一对与另一个酵母反式作用因子GRF2的结合序列同源的反向重复序列,以及包含CATCC基序的直接重复序列。缺失分析表明,这些元件也可作为转录调控元件发挥作用。

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