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Contribution of downstream promoter elements to transcriptional regulation of the rice tungro bacilliform virus promoter

机译:下游启动子元件对水稻通哥杆菌病毒启动子转录调控的贡献

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摘要

Downstream sequences influence activity of the rice tungro bacilliform virus (RTBV) promoter in protoplasts derived from cultured rice cells. We previously identified a DNA element located between positions +50 and +90 relative to the transcription start site to which rice nuclear proteins bind. In this study, using DNA UV crosslinking assays, we show that two rice nuclear proteins bind specifically to this DNA element. We demonstrate that the DNA element enhances RTBV promoter activity in a copy number-dependent manner when transferred to a position upstream of the promoter. In addition, using electro-phoretic mobility shift assays, we show that at least two novel nuclear proteins from rice cell suspension cultures bind to a subregion (from +50 to +59) of the DNA element and that a protein from rice root, but not shoot, nuclear extracts interacts with a perfect palindromic sequence motif located within the sequence +45 to +59. Furthermore, a position-dependent GAGA motif, present in three copies within downstream promoter sequences from +1 to +50, is involved in the regulation of RTBV promoter activity.
机译:下游序列影响源自培养的水稻细胞的原生质体中的水稻通哥杆菌病毒(RTBV)启动子的活性。我们先前确定了一个相对于水稻核蛋白结合的转录起始位点在+50和+90之间的DNA元件。在这项研究中,使用DNA UV交联测定法,我们显示了两个水稻核蛋白与该DNA元素特异性结合。我们证明,当转移到启动子上游位置时,DNA元件以拷贝数依赖性方式增强RTBV启动子活性。此外,使用电泳迁移率变动分析,我们显示了至少两种来自水稻细胞悬浮培养的新型核蛋白与DNA元件的一个子区域(从+50到+59)结合,并且来自稻根的一种蛋白质如果没有芽,核提取物与位于+45至+59序列中的完美回文序列基序相互作用。此外,存在于下游启动子序列中从+1至+50的三个副本中的位置依赖性GAGA基序参与RTBV启动子活性的调节。

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