Reversal of RNA Polymerase II Ubiquitylation by the Ubiquitin Protease Ubp3

Kristian Kvint; Jay P. Uhler; Michael J. Taschner; Stefan Sigurdsson; Hediye Erdjument-Bromage; Paul Tempst; Jesper Q. Svejstrup

doi:10.1016/j.molcel.2008.04.018

Reversal of RNA Polymerase II Ubiquitylation by the Ubiquitin Protease Ubp3

Kristian Kvint
, Jay P. Uhler
, Michael J. Taschner
, Stefan Sigurdsson
, Hediye Erdjument-Bromage
, Paul Tempst
, Jesper Q. Svejstrup

Faculty of Medicine

University of Iceland

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

Abstract

The final outcome of protein polyubiquitylation is often proteasome-mediated proteolysis, meaning that "proofreading" of ubiquitylation by ubiquitin proteases (UBPs) is crucial. Transcriptional arrest can trigger ubiquitin-mediated proteolysis of RNA polymerase II (RNAPII) so a UBP reversing RNAPII ubiquitylation might be expected. Here, we show that Ubp3 deubiquitylates RNAPII in yeast. Genetic characterization of ubp3 cells is consistent with a role in elongation, and Ubp3 can be purified with RNAPII, Def1, and the elongation factors Spt5 and TFIIF. This Ubp3 complex deubiquitylates both mono- and polyubiquitylated RNAPII in vitro, and ubp3 cells have elevated levels of ubiquitylated RNAPII in vivo. Moreover, RNAPII is degraded faster in a ubp3 mutant after UV irradiation. Problems posed by damage-arrested RNAPII are thought to be resolved either by removing the damage or degrading the polymerase. In agreement with this, cells with compromised DNA repair are better equipped to survive UV damage when UPB3 is deleted.

Original language	English
Pages (from-to)	498-506
Number of pages	9
Journal	Molecular Cell
Volume	30
Issue number	4
DOIs	https://doi.org/10.1016/j.molcel.2008.04.018
Publication status	Published - 23 May 2008

Bibliographical note

Funding Information: Peter Verrijzer is thanked for comments on the manuscript. This work was supported by a postdoctoral fellowship from the Swedish Research Council (to K.K.), by an EMBO long-term postdoctoral fellowship (to S.S.), and by grants from Cancer Research UK, AICR, and the EU (to J.Q.S.). We thank Catherine Dargemont for plasmids expressing point-mutated Ubp3.

Other keywords

DNA
PROTEINS

Access to Document

10.1016/j.molcel.2008.04.018

Cite this

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title = "Reversal of RNA Polymerase II Ubiquitylation by the Ubiquitin Protease Ubp3",

abstract = "The final outcome of protein polyubiquitylation is often proteasome-mediated proteolysis, meaning that {"}proofreading{"} of ubiquitylation by ubiquitin proteases (UBPs) is crucial. Transcriptional arrest can trigger ubiquitin-mediated proteolysis of RNA polymerase II (RNAPII) so a UBP reversing RNAPII ubiquitylation might be expected. Here, we show that Ubp3 deubiquitylates RNAPII in yeast. Genetic characterization of ubp3 cells is consistent with a role in elongation, and Ubp3 can be purified with RNAPII, Def1, and the elongation factors Spt5 and TFIIF. This Ubp3 complex deubiquitylates both mono- and polyubiquitylated RNAPII in vitro, and ubp3 cells have elevated levels of ubiquitylated RNAPII in vivo. Moreover, RNAPII is degraded faster in a ubp3 mutant after UV irradiation. Problems posed by damage-arrested RNAPII are thought to be resolved either by removing the damage or degrading the polymerase. In agreement with this, cells with compromised DNA repair are better equipped to survive UV damage when UPB3 is deleted.",

keywords = "DNA, PROTEINS",

author = "Kristian Kvint and Uhler, \{Jay P.\} and Taschner, \{Michael J.\} and Stefan Sigurdsson and Hediye Erdjument-Bromage and Paul Tempst and Svejstrup, \{Jesper Q.\}",

note = "Funding Information: Peter Verrijzer is thanked for comments on the manuscript. This work was supported by a postdoctoral fellowship from the Swedish Research Council (to K.K.), by an EMBO long-term postdoctoral fellowship (to S.S.), and by grants from Cancer Research UK, AICR, and the EU (to J.Q.S.). We thank Catherine Dargemont for plasmids expressing point-mutated Ubp3.",

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AU - Kvint, Kristian

AU - Uhler, Jay P.

AU - Taschner, Michael J.

AU - Sigurdsson, Stefan

AU - Erdjument-Bromage, Hediye

AU - Tempst, Paul

AU - Svejstrup, Jesper Q.

N1 - Funding Information: Peter Verrijzer is thanked for comments on the manuscript. This work was supported by a postdoctoral fellowship from the Swedish Research Council (to K.K.), by an EMBO long-term postdoctoral fellowship (to S.S.), and by grants from Cancer Research UK, AICR, and the EU (to J.Q.S.). We thank Catherine Dargemont for plasmids expressing point-mutated Ubp3.

PY - 2008/5/23

Y1 - 2008/5/23

N2 - The final outcome of protein polyubiquitylation is often proteasome-mediated proteolysis, meaning that "proofreading" of ubiquitylation by ubiquitin proteases (UBPs) is crucial. Transcriptional arrest can trigger ubiquitin-mediated proteolysis of RNA polymerase II (RNAPII) so a UBP reversing RNAPII ubiquitylation might be expected. Here, we show that Ubp3 deubiquitylates RNAPII in yeast. Genetic characterization of ubp3 cells is consistent with a role in elongation, and Ubp3 can be purified with RNAPII, Def1, and the elongation factors Spt5 and TFIIF. This Ubp3 complex deubiquitylates both mono- and polyubiquitylated RNAPII in vitro, and ubp3 cells have elevated levels of ubiquitylated RNAPII in vivo. Moreover, RNAPII is degraded faster in a ubp3 mutant after UV irradiation. Problems posed by damage-arrested RNAPII are thought to be resolved either by removing the damage or degrading the polymerase. In agreement with this, cells with compromised DNA repair are better equipped to survive UV damage when UPB3 is deleted.

AB - The final outcome of protein polyubiquitylation is often proteasome-mediated proteolysis, meaning that "proofreading" of ubiquitylation by ubiquitin proteases (UBPs) is crucial. Transcriptional arrest can trigger ubiquitin-mediated proteolysis of RNA polymerase II (RNAPII) so a UBP reversing RNAPII ubiquitylation might be expected. Here, we show that Ubp3 deubiquitylates RNAPII in yeast. Genetic characterization of ubp3 cells is consistent with a role in elongation, and Ubp3 can be purified with RNAPII, Def1, and the elongation factors Spt5 and TFIIF. This Ubp3 complex deubiquitylates both mono- and polyubiquitylated RNAPII in vitro, and ubp3 cells have elevated levels of ubiquitylated RNAPII in vivo. Moreover, RNAPII is degraded faster in a ubp3 mutant after UV irradiation. Problems posed by damage-arrested RNAPII are thought to be resolved either by removing the damage or degrading the polymerase. In agreement with this, cells with compromised DNA repair are better equipped to survive UV damage when UPB3 is deleted.

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KW - PROTEINS

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DO - 10.1016/j.molcel.2008.04.018

M3 - Article

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EP - 506

JO - Molecular Cell

JF - Molecular Cell

IS - 4

ER -

Reversal of RNA Polymerase II Ubiquitylation by the Ubiquitin Protease Ubp3

Abstract

Bibliographical note

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