Fischl Harry, Howe Françoise S, Furger Andre, Mellor Jane
Department of Biochemistry, University of Oxford, South Parks Road, Oxford OX1 3QU, UK.
Department of Biochemistry, University of Oxford, South Parks Road, Oxford OX1 3QU, UK.
Mol Cell. 2017 Feb 16;65(4):685-698.e8. doi: 10.1016/j.molcel.2017.01.006. Epub 2017 Feb 9.
RNA polymerase II (Pol2) movement through chromatin and the co-transcriptional processing and fate of nascent transcripts is coordinated by transcription elongation factors (TEFs) such as polymerase-associated factor 1 (Paf1), but it is not known whether TEFs have gene-specific functions. Using strand-specific nucleotide resolution techniques, we show that levels of Paf1 on Pol2 vary between genes, are controlled dynamically by environmental factors via promoters, and reflect levels of processing and export factors on the encoded transcript. High levels of Paf1 on Pol2 promote transcript nuclear export, whereas low levels reflect nuclear retention. Strains lacking Paf1 show marked elongation defects, although low levels of Paf1 on Pol2 are sufficient for transcription elongation. Our findings support distinct Paf1 functions: a core general function in transcription elongation, satisfied by the lowest Paf1 levels, and a regulatory function in determining differential transcript fate by varying the level of Paf1 on Pol2.
RNA聚合酶II(Pol2)在染色质中的移动以及新生转录本的共转录加工和命运,是由诸如聚合酶相关因子1(Paf1)等转录延伸因子(TEF)协调的,但目前尚不清楚TEF是否具有基因特异性功能。使用链特异性核苷酸分辨率技术,我们发现Pol2上Paf1的水平在不同基因之间存在差异,受环境因子通过启动子动态调控,并反映编码转录本上加工和输出因子的水平。Pol2上高水平的Paf1促进转录本的核输出,而低水平则反映核滞留。缺乏Paf1的菌株表现出明显的延伸缺陷,尽管Pol2上低水平的Paf1足以进行转录延伸。我们的研究结果支持Paf1的不同功能:在转录延伸中具有核心通用功能,由最低水平的Paf1实现;以及通过改变Pol2上Paf1的水平来决定转录本不同命运的调节功能。