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Guy G. Poirier
Ph.D.
Groupe de recherche sur les m茅canismes de poly(ADP-ribosyl)ation (Axe Cancer, CHUQ - Pavillon CHUL) Chaire du Canada en Prot茅omique
Plateforme prot茅omique du Centre de G茅nomique de Qu茅bec

鈥淩ole of poly(ADP-ribose)-binding proteins in DNA damage signalling and repair鈥�

One of the earliest step in the DNA damage signaling is the recruitment of poly(ADP-ribose) polymerase-1 (PARP-1) at the vicinity of the DNA strand break (DSB). In presence of DNA lesions, PARP-1 is rapidly activated and signals the presence of damage by attaching ADP-ribose units to chromatin-associated proteins. Poly(ADP-ribosyl)ation may function to organize the recruitment of repair proteins to the DSB by noncovalent pADPr-binding interactions. The central role of poly(ADP-ribose) (pADPr) in the recruitment and assembly of multiprotein repair complexes strenghten the role of DNA-dependent PARPs as regulators of genome maintenance pathways.

In spite of the importance of pADPr in the molecular basis of DNA damage response, few pADPr-binding proteins and pADPr-containing multiprotein complexes have been identified so far. Our laboratory is presently engaged in proteome-wide identification of proteins with affinity to pADPr. In silico prediction of pADPr-binding proteins, large-scale mass spectrometry-based proteome analysis and protein microarray analysis of pADPr-binding proteins were used to establish a comprehensive repertoire of pADPr-associated proteins. The involvement of putative pADPr-binding proteins in DNA damage signaling is presently under investigation using in vivo laser microirradiation-induced DNA damage which permits to evaluate the pADPr-dependent recruitment of protein at sites of DNA damage. We are currently using Multiple Reaction Monitoring (MRM), one of the most selective and sensitive mass spectrometry scan function, for the quantitative analysis of pADPr-containing protein complexes involved in DNA damage response and repair. In addition, we perform SILAC analysis (Stable Isotope Labeling with Amino acids in Cell culture) which allows quantitative assessment of proteins levels in anti-pADPr immunoprecipitation extracts with correlation with pADPr levels.

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Supported by grants from Canadian Institutes of Health Research (CIHR), the National Institutes of Health (NIH) and the Alberta Cancer Board.

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