Department of Molecular Cellular and Developmental Biology, Yale University, 219 Prospect Street, New Haven, CT 06511, USA; Systems Biology Institute, Yale University, 850 West Campus Drive, West Haven, CT 06516, USA.
Systems Biology Institute, Yale University, 850 West Campus Drive, West Haven, CT 06516, USA; Interdepartmental Program in Computational Biology and Bioinformatics, Yale University, 300 George Street, Suite 501, New Haven, CT 06511, USA.
Cell Rep. 2018 Oct 16;25(3):737-748.e4. doi: 10.1016/j.celrep.2018.09.050.
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.
尽管在理解启动子结构对转录活性的影响方面已经取得了进展，但对于染色质结构在转录中的作用知之甚少。之前的工作假设，双向 GAL1/GAL10 启动子的转录是通过其 UAS 区域围绕非标准核小体进行环化来控制的。在这里，通过在高分辨率下编辑 GAL1/GAL10 启动子，我们深入了解了双向表达控制。我们证明，UAS 内的第一个和第四个 Gal4 结合位点对启动子激活没有功能贡献。相反，这些位点以及附近的调控区域，有助于基因表达的定向调控。此外，Gal4 与第三个结合位点的结合对于基因表达至关重要，而与其他三个位点的结合不足以激活转录。由于 GAL1/GAL10 UAS 可以在许多真核生物中激活基因表达，因此预计所提出的调控机制将在真核生物进化枝中广泛发挥作用。
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