Investigation of PARP-1, PARP-2, and PARG interactomes by affinity-purification mass spectrometry

Poly(ADP-ribose) polymerases (PARPs) catalyze the formation of poly(ADP-ribose) (pADPr), a post-translational modification involved in several important biological processes, namely surveillance of genome integrity, cell cycle progression, initiation of the DNA damage response, apoptosis, and regula...

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Bibliographic Details
Published in:Proteome science 2010-04, Vol.8 (1), p.22-22, Article 22
Main Authors: Isabelle, Maxim, Moreel, Xavier, Gagné, Jean-Philippe, Rouleau, Michèle, Ethier, Chantal, Gagné, Pierre, Hendzel, Michael J, Poirier, Guy G
Format: Article
Language:English
Subjects:
Apoptosis
Cell cycle
DNA
DNA damage
DNA repair
Enzymes
Genomes
Glucose metabolism
Health aspects
Medical research
Ontology
Probability
Proteins
Proteomics
Reorganization and restructuring
RNA
Software
Spectrum analysis
Statistical analysis
Trypsin
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Summary:Poly(ADP-ribose) polymerases (PARPs) catalyze the formation of poly(ADP-ribose) (pADPr), a post-translational modification involved in several important biological processes, namely surveillance of genome integrity, cell cycle progression, initiation of the DNA damage response, apoptosis, and regulation of transcription. Poly(ADP-ribose) glycohydrolase (PARG), on the other hand, catabolizes pADPr and thereby accounts for the transient nature of poly(ADP-ribosyl)ation. Our investigation of the interactomes of PARP-1, PARP-2, and PARG by affinity-purification mass spectrometry (AP-MS) aimed, on the one hand, to confirm current knowledge on these interactomes and, on the other hand, to discover new protein partners which could offer insights into PARPs and PARG functions. PARP-1, PARP-2, and PARG were immunoprecipitated from human cells, and pulled-down proteins were separated by gel electrophoresis prior to in-gel trypsin digestion. Peptides were identified by tandem mass spectrometry. Our AP-MS experiments resulted in the identifications of 179 interactions, 139 of which are novel interactions. Gene Ontology analysis of the identified protein interactors points to five biological processes in which PARP-1, PARP-2 and PARG may be involved: RNA metabolism for PARP-1, PARP-2 and PARG; DNA repair and apoptosis for PARP-1 and PARP-2; and glycolysis and cell cycle for PARP-1. This study reveals several novel protein partners for PARP-1, PARP-2 and PARG. It provides a global view of the interactomes of these proteins as well as a roadmap to establish the systems biology of poly(ADP-ribose) metabolism.
ISSN:1477-5956
1477-5956
DOI:10.1186/1477-5956-8-22