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Mammalian NET-seq reveals genome-wide nascent transcription coupled to RNA processing

GSE60358 Homo sapiens Expression profiling by high throughput sequencing; Other 28 samples Submitted 2015/04/23 Platform GPL16791Platform GPL11154
Summary
We have generated single-nucleotide resolution, nascent transcription profiles from HeLa cells by developing Native Elongation Transcript sequencing technology for mammalian chromatin (mNET-seq). Our extensive data sets provide a substantial resource to study mammalian nascent transcript profiles. We reveal unanticipated phosphorylation states for RNA polymerase II C-terminal domain (Pol II CTD) at both gene ends. We also observe that following 5’ splice site cleavage by the spliceosome, upstream exon transcripts are tethered to Pol II CTD phosphorylated on the serine 5 position (S5P) which is accumulated over downstream exons. We further show that depletion of termination factors substantially reduces Pol II pausing at gene ends leading to termination defects. Remarkably termination factors play an additional promoter role by restricting non-productive RNA synthesis and redistributing Pol II CTD S2P to promoters. These data demonstrate that CTD phosphorylation is more dynamic and variably distributed across mammalian transcription units than previously envisaged.
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Also filed as BioProject PRJNA258089 and SRA study SRP045447. Searching any of these in the dataset finder brings you back here.

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