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Elife. 2017 Oct 13;6. pii: e27082. doi: 10.7554/eLife.27082.

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

Author information

1
Wellcome Trust Centre for Cell Biology, University of Edinburgh, Edinburgh, Scotland.
2
Friedrich Miescher Institute for Biomedical Research, Basel, Switzerland.
3
Institute of Biotechnology, Technische Universität Berlin, Berlin, Germany.
4
MRC Human Genetics Unit, Institute of Genetics and Molecular Medicine, University of Edinburgh, Edinburgh, Scotland.

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.

KEYWORDS:

RNA polymerase; RNA processing; S. cerevisiae; UV crosslinking; chromosomes; genes; protein modification; transcription; ubiquitin protease

PMID:
29027900
PMCID:
PMC5673307
DOI:
10.7554/eLife.27082
[Indexed for MEDLINE]
Free PMC Article

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