RNA polymerase II activity revealed by GRO-seq and pNET-seq in Arabidopsis
RNA polymerase II activity revealed by GRO-seq and pNET-seq in Arabidopsis
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GRO-seq 和 pNET-seq 揭示拟南芥中 RNA 聚合酶 II 活性
DOI：
10.1038/s41477-018-0280-0
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发表时间：
2018-12-01
期刊：
影响因子：
18
通讯作者：
Dong, Zhicheng
中科院分区：
文献类型：
--
作者：
Zhu, Jiafu;Liu, Min;Dong, Zhicheng
RNA聚合酶II（Pol II）在基因表达中起重要作用。我们使用植物天然延伸转录测序和全局连续测序来分析拟南芥中全基因组的新生RNA。我们发现Pol II倾向于在转录起始位点（TSS）的下游积累。此外，具有未磷酸化羧基末端结构域（CTD）的Pol II主要在TSS下游积累，而具有Ser 5P CTD的Pol II与剪接体缔合，具有Ser 2P CTD的Pol II在多聚腺苷酸化位点（PAS）下游250个碱基对内呈现尖锐峰。Pol II在启动子近端区域和PAS后暂停影响转录速率。有趣的是，活性基因可以根据不同的转录模式分为三个簇。我们证明，这两种方法是适合研究Pol II动力学在植物。虽然转录在真核生物中总体上是保守的，但存在植物特异性调节。
RNA polymerase II (Pol II) plays an essential role in gene expression. We used plant native elongating transcript sequencing and global run-on sequencing to profile nascent RNAs genome wide in Arabidopsis. We found that Pol II tends to accumulate downstream of the transcription start site (TSS). Moreover, Pol II with an unphosphorylated carboxyl-terminal domain (CTD) mainly accumulates downstream of the TSS, while Pol II with a Ser 5P CTD associates with spliceosomes, and Pol II with a Ser 2P CTD presents a sharp peak within 250 base pairs downstream of the polyadenylation site (PAS). Pol II pausing both at promoter-proximal regions and after PAS affects the transcription rate. Interestingly, active genes can be classified into three clusters based on the different modes of transcription. We demonstrate that these two methods are suitable to study Pol II dynamics in planta. Although transcription is conserved overall within eukaryotes, there is plant-specific regulation.