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

作者：Maxim Isabelle, Xavier Moreel, Jean‐Philippe Gagné, Michèle Rouleau, Chantal Éthier, Pierre Gagné, Michael J. Hendzel, Guy G. Poirier · 发表于：Proteome Science · 年份：2010 · DOI：10.1186/1477-5956-8-22 · 被引用次数：163 · 研究领域：PARP inhibition in cancer therapy、RNA Research and Splicing、Clusterin in disease pathology

BACKGROUND: 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. RESULTS: 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. CONCLUSIONS: This study reveals several novel protein partners for PARP-1, PARP-2 and P...