RNA polymerase II CTD is dispensable for transcription and required for termination in human cells

Yousra Yahia, Alexia Pigeot, Amal Zine El Aabidine, Nilay Shah, Nezih Karasu, Ignasi Forné, Stefan Krebs, Helmut Blum, Cyril Esnault, Tom Sexton, Axel Imhof, Dirk Eick, Jean Christophe Andrau

Research output: Contribution to journalArticlepeer-review

3 Scopus citations

Abstract

The largest subunit of RNA polymerase (Pol) II harbors an evolutionarily conserved C-terminal domain (CTD), composed of heptapeptide repeats, central to the transcriptional process. Here, we analyze the transcriptional phenotypes of a CTD-Δ5 mutant that carries a large CTD truncation in human cells. Our data show that this mutant can transcribe genes in living cells but displays a pervasive phenotype with impaired termination, similar to but more severe than previously characterized mutations of CTD tyrosine residues. The CTD-Δ5 mutant does not interact with the Mediator and Integrator complexes involved in the activation of transcription and processing of RNAs. Examination of long-distance interactions and CTCF-binding patterns in CTD-Δ5 mutant cells reveals no changes in TAD domains or borders. Our data demonstrate that the CTD is largely dispensable for the act of transcription in living cells. We propose a model in which CTD-depleted Pol II has a lower entry rate onto DNA but becomes pervasive once engaged in transcription, resulting in a defect in termination.

Original languageEnglish
Article numbere56150
JournalEMBO Reports
Volume24
Issue number9
DOIs
StatePublished - 6 Sep 2023
Externally publishedYes

Keywords

  • Pol II interactome
  • RNA polymerase II (Pol II)
  • carboxy-terminal domain (CTD)
  • mammalian transcription
  • termination

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