Shetty Ameet, Kallgren Scott P, Demel Carina, Maier Kerstin C, Spatt Dan, Alver Burak H, Cramer Patrick, Park Peter J, Winston Fred
Department of Genetics, Harvard Medical School, Boston, MA 02115, USA.
Department of Biomedical Informatics, Harvard Medical School, Boston, MA 02115, USA.
Mol Cell. 2017 Apr 6;66(1):77-88.e5. doi: 10.1016/j.molcel.2017.02.023. Epub 2017 Mar 30.
Spt5 is an essential and conserved factor that functions in transcription and co-transcriptional processes. However, many aspects of the requirement for Spt5 in transcription are poorly understood. We have analyzed the consequences of Spt5 depletion in Schizosaccharomyces pombe using four genome-wide approaches. Our results demonstrate that Spt5 is crucial for a normal rate of RNA synthesis and distribution of RNAPII over transcription units. In the absence of Spt5, RNAPII localization changes dramatically, with reduced levels and a relative accumulation over the first ∼500 bp, suggesting that Spt5 is required for transcription past a barrier. Spt5 depletion also results in widespread antisense transcription initiating within this barrier region. Deletions of this region alter the distribution of RNAPII on the sense strand, suggesting that the barrier observed after Spt5 depletion is normally a site at which Spt5 stimulates elongation. Our results reveal a global requirement for Spt5 in transcription elongation.
Spt5是一种在转录和共转录过程中发挥作用的必需且保守的因子。然而,对于转录过程中Spt5需求的许多方面,我们了解甚少。我们使用四种全基因组方法分析了粟酒裂殖酵母中Spt5缺失的后果。我们的结果表明,Spt5对于正常的RNA合成速率以及RNA聚合酶II(RNAPII)在转录单元上的分布至关重要。在没有Spt5的情况下,RNAPII的定位发生显著变化,水平降低且在最初约500个碱基对处相对积累,这表明Spt5是越过一个障碍进行转录所必需的。Spt5缺失还导致在这个障碍区域内广泛启动反义转录。该区域的缺失改变了有义链上RNAPII的分布,这表明在Spt5缺失后观察到的障碍通常是Spt5刺激延伸的位点。我们的结果揭示了转录延伸过程中对Spt5的全局需求。