TY - JOUR TI - RNA polymerase II stalling at pre-mRNA splice sites is enforced by ubiquitination of the catalytic subunit AU - Milligan, Laura AU - Sayou, Camille AU - Tuck, Alex AU - Auchynnikava, Tatsiana AU - Reid, Jane EA AU - Alexander, Ross AU - Alves, Flavia de Lima AU - Allshire, Robin AU - Spanos, Christos AU - Rappsilber, Juri AU - Beggs, Jean D AU - Kudla, Grzegorz AU - Tollervey, David A2 - Proudfoot, Nick J VL - 6 PY - 2017 DA - 2017/10/13 SP - e27082 C1 - eLife 2017;6:e27082 DO - 10.7554/eLife.27082 UR - https://doi.org/10.7554/eLife.27082 AB - 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. KW - RNA processing KW - UV crosslinking KW - transcription KW - RNA polymerase KW - ubiquitin protease KW - protein modification JF - eLife SN - 2050-084X PB - eLife Sciences Publications, Ltd ER -