Proteome-wide identification of poly(ADP-Ribosyl)ation targets in different genotoxic stress responses

Poly(ADP-ribos)ylation (PARylation) is a reversible posttranslational modification found in higher eukaryotes. However, little is known about PARylation acceptor proteins. Here, we describe a sensitive proteomics approach based on high-accuracy quantitative mass spectrometry for the identification o...

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Bibliographic Details
Published in:Molecular cell Vol. 52; no. 2; p. 272
Main Authors: Jungmichel, Stephanie, Rosenthal, Florian, Altmeyer, Matthias, Lukas, Jiri, Hottiger, Michael O, Nielsen, Michael L
Format: Journal Article
Language:English
Published: United States 24.10.2013
Subjects:
Antineoplastic Agents, Alkylating - pharmacology
Cell Line, Tumor
Cell Nucleus - genetics
Cell Nucleus - metabolism
Chromatography, Liquid
DNA Damage
DNA-Binding Proteins - genetics
DNA-Binding Proteins - metabolism
Electrophoresis, Polyacrylamide Gel
Green Fluorescent Proteins - genetics
Green Fluorescent Proteins - metabolism
HeLa Cells
Humans
Hydrogen Peroxide - pharmacology
Methyl Methanesulfonate - pharmacology
Microscopy, Confocal
Nuclear Proteins - genetics
Nuclear Proteins - metabolism
Oxidants - pharmacology
Poly Adenosine Diphosphate Ribose - metabolism
Protein Processing, Post-Translational - drug effects
Protein Processing, Post-Translational - radiation effects
Proteome - genetics
Proteome - metabolism
Proteomics - methods
Radiation, Ionizing
RNA Interference
Signal Transduction - drug effects
Signal Transduction - genetics
Signal Transduction - radiation effects
Tandem Mass Spectrometry
TATA-Binding Protein Associated Factors - genetics
TATA-Binding Protein Associated Factors - metabolism
Transcription Factors - genetics
Transcription Factors - metabolism
ISSN:1097-4164, 1097-4164
Online Access:Get more information
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Summary:Poly(ADP-ribos)ylation (PARylation) is a reversible posttranslational modification found in higher eukaryotes. However, little is known about PARylation acceptor proteins. Here, we describe a sensitive proteomics approach based on high-accuracy quantitative mass spectrometry for the identification of PARylated proteins induced under different cellular stress conditions. While confirming the majority of known PARylated substrates, our screen identifies numerous additional PARylation targets. In vivo and in vitro validation of acceptor proteins confirms that our methodology targets covalent PARylation. Nuclear proteins encompassing nucleic acid binding properties are prominently PARylated upon genotoxic stress, consistent with the nuclear localization of ARTD1/PARP1 and ARTD2/PARP2. Distinct differences in proteins becoming PARylated upon various genotoxic insults are observed, exemplified by the PARylation of RNA-processing factors THRAP3 and TAF15 under oxidative stress. High-content imaging reveals that PARylation affects the nuclear relocalization of THRAP3 and TAF15, demonstrating the potential of our approach to uncover hitherto unappreciated processes being controlled by specific genotoxic-stress-induced PARylation.
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ISSN:1097-4164
1097-4164
DOI:10.1016/j.molcel.2013.08.026