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

doi:10.7554/eLife.27082

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

Research output: Contribution to journal › Article › peer-review

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 language	English
Article number	e27082
Number of pages	27
Journal	eLIFE
Volume	6
DOIs	https://doi.org/10.7554/eLife.27082
Publication status	Published - 13 Oct 2017

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10.7554/eLife.27082Licence: Creative Commons: Attribution (CC-BY)

TollerveyEtaleLife2020RNAPolymeraseIIStallingAtPre-mRNASpliceSitesIsEnforcedByUbiquitinationFinal published version, 1.98 MBLicence: Creative Commons: Attribution (CC-BY)

FUNCTIONAL CONSEQUENCES OF SYNONOMOUS MUTATIONS IN EUKARYOTIC CELLS
Kudla, G. (Principal Investigator)
UK-based charities
1/08/12 → 31/01/18
Project: Research
HGU Core Award Apr 12 - MAr 18
Hastie, N. (Principal Investigator), Adams, I. (Co-investigator), Baldock, R. (Co-investigator), Bickmore, W. (Co-investigator), Caceres, J. (Co-investigator), Dorin, J. (Co-investigator), FitzPatrick, D. (Co-investigator), Haley, C. (Co-investigator), Hill, B. (Co-investigator), Jackson, I. (Co-investigator), Jackson, A. (Co-investigator), Kudla, G. (Co-investigator), Meehan, R. (Co-investigator) & Patton, E. (Co-investigator)
MRC
1/04/12 → 31/03/18
Project: Research

David Tollervey
- School of Biological Sciences - Professor of RNA Biology
- Centre for Engineering Biology
Person: Academic: Research Active

Cite this

@article{d24ac7540e3f4bb1b01fc0e752004e20,

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

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.",

author = "Laura Milligan and Camille Sayou and Alex Tuck and Tatsiana Auchynnikava and Reid, {Jane E A} and Ross Alexander and Alves, {Flavia De Lima} and Robin Allshire and Christos Spanos and Juri Rappsilber and Beggs, {Jean D} and Grzegorz Kudla and David Tollervey",

year = "2017",

month = oct,

day = "13",

doi = "10.7554/eLife.27082",

language = "English",

volume = "6",

journal = "eLIFE",

issn = "2050-084X",

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T1 - 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 E A

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

PY - 2017/10/13

Y1 - 2017/10/13

N2 - 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.

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.

U2 - 10.7554/eLife.27082

DO - 10.7554/eLife.27082

M3 - Article

SN - 2050-084X

VL - 6

JO - eLIFE

JF - eLIFE

M1 - e27082

ER -

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

Abstract

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Projects

FUNCTIONAL CONSEQUENCES OF SYNONOMOUS MUTATIONS IN EUKARYOTIC CELLS

HGU Core Award Apr 12 - MAr 18

Profiles

David Tollervey

Cite this