Insights into Bidirectional Gene Expression Control Using the Canonical GAL1/GAL10 Promoter.
Insights into Bidirectional Gene Expression Control Using the Canonical GAL1/GAL10 Promoter.
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DOI：
10.1016/j.celrep.2018.09.050
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发表时间：
2018-10-16
期刊：
影响因子：
8.8
通讯作者：
Acar M
中科院分区：
文献类型：
--
作者：
Elison GL;Xue Y;Song R;Acar M
尽管在理解启动子结构对转录活性的影响方面取得了进展，但关于染色质结构在转录中所起的作用的知识有限。以前的工作假设，从双向GAL 1/GAL 10启动子的转录控制通过其UAS区域周围的非标准核小体的循环。在这里，通过以高分辨率编辑GAL 1/GAL 10启动子，我们提供了对双向表达控制的见解。我们证明，第一和第四Gal 4结合位点内的UAS不功能上有助于启动子激活。相反，这些位点，沿着与附近的调控区，有助于基因表达的定向调控。此外，Gal 4结合到第三个结合位点对基因表达是至关重要的，而结合到其他三个位点不足以激活转录。由于GAL 1/GAL 10 UAS可以激活许多真核生物中的基因表达，因此预期所提出的调控机制将在真核进化枝中广泛发挥作用。Elison等证明了从经典GAL 1/GAL 10启动子的转录是如何双向控制的。使用CRISPR编辑启动子内容，他们发现了对定向基因表达控制至关重要的不同调控位点。他们阐明了四个Gal 4结合位点在影响转录激活和方向性方面的差异性。
Despite advances made in understanding the effects of promoter structure on transcriptional activity, limited knowledge exists regarding the role played by chromatin architecture in transcription. Previous work hypothesized that transcription from the bidirectional GAL1/GAL10 promoter is controlled through looping of its UAS region around a nonstandard nucleosome. Here, by editing the GAL1/GAL10 promoter at high resolution, we provide insights into bidirectional expression control. We demonstrate that the first and fourth Gal4 binding sites within the UAS do not functionally contribute to promoter activation. Instead, these sites, along with nearby regulatory regions, contribute to the directional regulation of gene expression. Furthermore, Gal4 binding to the third binding site is critical for gene expression, while binding to the other three sites is not sufficient for transcriptional activation. Because the GAL1/GAL10 UAS can activate gene expression in many eukaryotes, the regulatory mechanism presented is expected to operate broadly across the eukaryotic clade. Elison et al. demonstrate how transcription from the canonical GAL1/GAL10 promoter is controlled bidirectionally. Editing the promoter content using CRISPR, they uncover distinct regulatory sites that are essential for directional gene expression control. They elucidate the differential nature of the four Gal4 binding sites in affecting transcriptional activation and directionality.
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