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

• Published in: eLife Vol. 6
• Main Authors: Milligan, Laura, Sayou, Camille, Tuck, Alex, Auchynnikava, Tatsiana, Reid, Jane Ea, Alexander, Ross, Alves, Flavia de Lima, Allshire, Robin, Spanos, Christos, Rappsilber, Juri, Beggs, Jean D, Kudla, Grzegorz, Tollervey, David
• Format: Journal Article
• Language: English
• Published: England eLife Science Publications, Ltd 13.10.2017; eLife Sciences Publications Ltd; eLife Sciences Publication; eLife Sciences Publications, Ltd
• Subjects: Catalytic Domain; Chromosomes and Gene Expression; Deoxyribonucleic acid; DNA; DNA-directed RNA polymerase; Elongation; Endopeptidases - metabolism; Genomes; Health surveillance; Journalists; Life Sciences; Messenger RNA; Models, Biological; Mutant Proteins - genetics; Mutant Proteins - metabolism; Mutation, Missense; Poly(A); Proteases; protein modification; Proteins; RNA; RNA polymerase; RNA Polymerase II - genetics; RNA Polymerase II - metabolism; RNA Precursors - metabolism; RNA processing; Saccharomyces cerevisiae Proteins - metabolism; Spectrum analysis; Splicing; Standard deviation; Transcription; Transcription (Genetics); Ubiquitin; ubiquitin protease; Ubiquitination; UV crosslinking; Germany; transcription; UV crosslinking; genes; chromosomes; protein modification; RNA polymerase; RNA processing; S. cerevisiae; ubiquitin protease
• ISSN: 2050-084X; 2050-084X
• DOI: 10.7554/eLife.27082

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 K R 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.