GSE210601
RNA Polymerase II CTD is dispensable for transcription in vivo and required for termination
The largest subunit of RNA polymerase (Pol) II harbors a evolutionary conserved C-Terminal-Domain (CTD), composed of a repetition of heptapeptides, central in the transcriptional process. Here, we have analysed the transcriptional phenotypes of a CTD-?5 mutant that carries a large CTD truncation. Unexpectedly, our results indicate that this mutant can widely transcribe genes in living cells but displays an extreme pervasive phenotype with severely impaired termination, similar to but more severely impaired than previously characterized mutations of CTD tyrosine residues. The CTD-?5 mutant has also lost its interactions with the Mediator and Integrator complexes involved in activation of transcription and maturation of RNAs. Examination of long-distance interactions and CTCF binding pattern in CTD-?5 did not indicate major changes in TAD domains or borders. Our data demonstrate that the CTD is largely dispensable for the act of transcription per se in living cells. We propose a model in which CTD-depleted Pol II has a lower entry rate into DNA but becomes extremely pervasive once engaged in transcription, resulting in a global loss of termination. Overall design: This study was performed in a human Raji cell line. It contains ChIP-seq data for HA-tagged RNA Pol II, H3K4me1, H3K4me3 and CTCF as well as RNA-seq data for PolyA-RNA and chromatin-associated RNA (ChrRNA) and HiC data. All sequencing were performed in paired-end sequencing runs.