RNA polymerase II stalling at pre-mRNA splice sites is enforced by ubiquitination of the catalytic subunit

Laura Milligan, Camille Sayou, Alex Tuck, Tatsiana Auchynnikava, Jane E. A. Reid, Ross Alexander, Flavia de Lima Alves, Robin Allshire, Christos Spanos, Juri Rappsilber, Jean D. Beggs, Grzegorz Kudla, David Tollervey

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8 Citations (Scopus)
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Abstract

Numerous links exist between co-transcriptional RNA processing and the transcribing RNAPII. In particular, pre-mRNA splicing was reported to be associated with slowed RNAPII elongation. Here, we identify a site of ubiquitination (K1246) in the catalytic subunit of RNAPII close to the DNA entry path. Ubiquitination was increased in the absence of the Bre5-Ubp3 ubiquitin protease complex. Bre5 binds RNA in vivo, with a preference for exon 2 regions of intron-containing pre-mRNAs and poly(A) proximal sites. Ubiquitinated RNAPII showed similar enrichment. The absence of Bre5 led to impaired splicing and defects in RNAPII elongation in vivo on a splicing reporter construct. Strains expressing RNAPII with a K1246R mutation showed reduced co-transcriptional splicing. We propose that ubiquinitation of RNAPII is induced by RNA processing events and linked to transcriptional pausing, which is released by Bre5-Ubp3 associated with the nascent transcript.

Original languageEnglish
Article numbere27082
JournaleLife
Volume6
DOIs
Publication statusPublished - 13 Oct 2017

ASJC Scopus subject areas

  • Neuroscience(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

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    Milligan, L., Sayou, C., Tuck, A., Auchynnikava, T., Reid, J. E. A., Alexander, R., de Lima Alves, F., Allshire, R., Spanos, C., Rappsilber, J., Beggs, J. D., Kudla, G., & Tollervey, D. (2017). RNA polymerase II stalling at pre-mRNA splice sites is enforced by ubiquitination of the catalytic subunit. eLife, 6, [e27082]. https://doi.org/10.7554/eLife.27082