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几种不同的上游启动子元件可增强BPV-1 E2蛋白的反式激活作用。

Several different upstream promoter elements can potentiate transactivation by the BPV-1 E2 protein.

作者信息

Ham J, Dostatni N, Arnos F, Yaniv M

机构信息

Département des Biotechnologies, UA1149 du Centre National de la Recherche Scientifique, Institut Pasteur, Paris, France.

出版信息

EMBO J. 1991 Oct;10(10):2931-40. doi: 10.1002/j.1460-2075.1991.tb07843.x.

Abstract

The enhancer and upstream promoter regions of RNA polymerase II transcribed genes modulate the rate of transcription initiation and establish specific patterns of gene expression. Both types of region consist of clusters of DNA binding sites for nuclear proteins. To determine how efficiently the same factor can activate transcription when acting as an enhancer or promoter factor, we have studied transactivation by the BPV-1 E2 protein, a papillomavirus transcriptional regulator. By cotransfecting a BPV-1 E2 expression vector and a series of reporter plasmids containing well-defined chimeric promoters we have found that whilst E2 can strongly stimulate complex promoters such as that of the HSV tk gene, it does not efficiently activate constructions containing only a TATA box and initiation site. We show that insertion of upstream promoter elements, but not of spacer DNA, between E2 binding sites and the TATA box greatly increases E2 activation. This effect was observed with more than one type of upstream promoter element, is not related to the strength of the promoter and is unlikely to result from co-operative DNA binding by E2 and the transcription factors tested. These results would suggest that E2 has the properties of an enhancer rather than promoter factor and that in certain cases promoter and enhancer factors may affect different steps in the process of transcriptional activation.

摘要

RNA聚合酶II转录基因的增强子和上游启动子区域调节转录起始速率并建立特定的基因表达模式。这两种区域都由核蛋白的DNA结合位点簇组成。为了确定同一因子作为增强子或启动子因子时激活转录的效率,我们研究了乳头瘤病毒转录调节因子BPV-1 E2蛋白的反式激活作用。通过共转染BPV-1 E2表达载体和一系列含有明确嵌合启动子的报告质粒,我们发现虽然E2可以强烈刺激复杂启动子,如HSV tk基因的启动子,但它不能有效激活仅含有TATA盒和起始位点的构建体。我们表明,在E2结合位点和TATA盒之间插入上游启动子元件而非间隔DNA可大大增加E2的激活作用。用不止一种类型的上游启动子元件观察到了这种效应,它与启动子的强度无关,也不太可能是由E2与所测试的转录因子的协同DNA结合导致的。这些结果表明E2具有增强子而非启动子因子的特性,并且在某些情况下,启动子因子和增强子因子可能影响转录激活过程中的不同步骤。

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