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Yahia, Y.* ; Pigeot, A.* ; El Aabidine, A.Z.* ; Shah, N. ; Karasu, N.* ; Forne, I.* ; Krebs, S.* ; Blum, H.* ; Esnault, C.* ; Sexton, T.* ; Imhof, A.* ; Eick, D. ; Andrau, J.C.*

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

EMBO Rep. 24:e56150 (2023)

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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.

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