Department of Plant and Environmental Sciences, Copenhagen Plant Science Centre, University of Copenhagen, Frederiksberg, Denmark.
Department of Biochemistry and Biophysics, Texas A&M University, College Station, TX, USA.
EMBO Rep. 2020 Apr 3;21(4):e49315. doi: 10.15252/embr.201949315. Epub 2020 Feb 27.
RNA polymerase II (RNAPII) transcription is crucial for gene expression. RNAPII density peaks at gene boundaries, associating these key regions for gene expression control with limited RNAPII movement. The connections between RNAPII transcription speed and gene regulation in multicellular organisms are poorly understood. Here, we directly modulate RNAPII transcription speed by point mutations in the second largest subunit of RNAPII in Arabidopsis thaliana. A RNAPII mutation predicted to decelerate transcription is inviable, while accelerating RNAPII transcription confers phenotypes resembling auto-immunity. Nascent transcription profiling revealed that RNAPII complexes with accelerated transcription clear stalling sites at both gene ends, resulting in read-through transcription. The accelerated transcription mutant NRPB2-Y732F exhibits increased association with 5' splice site (5'SS) intermediates and enhanced splicing efficiency. Our findings highlight potential advantages of RNAPII stalling through local reduction in transcription speed to optimize gene expression for the development of multicellular organisms.
RNA 聚合酶 II(RNAPII)转录对于基因表达至关重要。RNAPII 在基因边界处出现密度峰值,将这些关键的基因表达调控区域与有限的 RNAPII 运动联系起来。在多细胞生物中,RNAPII 转录速度与基因调控之间的联系还知之甚少。在这里,我们通过在拟南芥的 RNAPII 第二大亚基上的点突变直接调节 RNAPII 转录速度。预测会减缓转录的 RNAPII 突变是不可存活的,而加速 RNAPII 转录赋予类似于自身免疫的表型。新生转录谱分析显示,具有加速转录的 RNAPII 复合物可以在基因两端清除stalling 位点,从而导致通读转录。加速转录的突变体 NRPB2-Y732F 与 5' 剪接位点(5'SS)中间体的结合增加,并提高了剪接效率。我们的发现强调了通过局部降低转录速度来优化多细胞生物发育过程中基因表达的 RNAPII stalling 的潜在优势。