Analysis of RNA polymerase II ubiquitylation and proteasomal degradation.
Degradation
RNA polymerase II
UV-irradiation
Ubiquitylation
Journal
Methods (San Diego, Calif.)
ISSN: 1095-9130
Titre abrégé: Methods
Pays: United States
ID NLM: 9426302
Informations de publication
Date de publication:
15 04 2019
15 04 2019
Historique:
received:
07
12
2018
revised:
04
02
2019
accepted:
06
02
2019
pubmed:
16
2
2019
medline:
24
6
2020
entrez:
16
2
2019
Statut:
ppublish
Résumé
Transcribing RNA polymerase II (RNAPII) is decorated by a plethora of post-translational modifications that mark different stages of transcription. One important modification is RNAPII ubiquitylation, which occurs in response to numerous different stimuli that cause RNAPII stalling, such as DNA damaging agents, RNAPII inhibitors, or depletion of the nucleotide pool. Stalled RNAPII triggers a so-called "last resort pathway", which involves RNAPII poly-ubiquitylation and proteasome-mediated degradation. Different approaches have been described to study RNAPII poly-ubiquitylation and degradation, each method with its own advantages and caveats. Here, we describe optimised strategies for detecting ubiquitylated RNAPII and studying its degradation, but these protocols are suitable for studying other ubiquitylated proteins as well.
Identifiants
pubmed: 30769100
pii: S1046-2023(18)30312-8
doi: 10.1016/j.ymeth.2019.02.005
pmc: PMC6617506
pii:
doi:
Substances chimiques
RNA Polymerase II
EC 2.7.7.-
Proteasome Endopeptidase Complex
EC 3.4.25.1
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
146-156Subventions
Organisme : Cancer Research UK
ID : FC001166
Pays : United Kingdom
Organisme : Wellcome Trust
ID : FC001166
Pays : United Kingdom
Organisme : Medical Research Council
ID : FC001166
Pays : United Kingdom
Informations de copyright
Copyright © 2019 The Authors. Published by Elsevier Inc. All rights reserved.
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