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Decoding the DNA damage signalling in C. elegans by proteomic analyses of ADP-ribosylation

Objectif

Preservation of genome integrity and stability is critical for survival and propagation of individuals and species. Organisms have thus evolved rapid and efficient mechanisms -collectively termed the DNA damage response (DDR)- to combat threats posed by DNA damage. Among these, the post-translational modification (PTM) ADP-ribosylation (ADPR) plays a decisive role in effective DDR. Although much is known about the relevance of ADPR upon DNA injury, the underlying molecular mechanisms are still poorly understood and no systematic, unbiased proteome-wide study to determine ADPR targets in vivo has been conducted to date. The most challenging and innovative goal of this proposal is to profile in vivo ADP-ribosylated peptides during DDR in the well-established model system Caenorhabditis elegans by the combined application of advanced proteomic approaches. I will develop novel enrichment strategies that will allow me to confidently map for the first time all the sites of ADPR throughout the C. elegans proteome. I will take advantage of SILAN technology, which allows stable isotope labelling in C. elegans, to determine the quantitative profiles of thousands of ADPR sites during DDR. By bioinformatic analysis, I will select candidate proteins for further investigation of the biological role of DNA damage-induced ADPR. I will then test the biological impact of ADPR of these candidates through site-specific mutation of their identified modification sites. Finally, I will perform biochemical, molecular and functional experiments to study how ADPR alters the activity of these proteins and thereby characterize the mode of ADPR action. Understanding the regulatory networks that underlie such a complex biological process at the organism level will provide new insights for improved treatment of DNA damage-related diseases including cancer. This is an ambitious, innovative, cross-disciplinary project at the forefront of two exciting fields, biology and proteomics.

Coordinateur

MAX-PLANCK-GESELLSCHAFT ZUR FORDERUNG DER WISSENSCHAFTEN EV
Contribution nette de l'UE
€ 159 460,80
Adresse
HOFGARTENSTRASSE 8
80539 Munchen
Allemagne

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Région
Bayern Oberbayern München, Kreisfreie Stadt
Type d’activité
Research Organisations
Liens
Coût total
€ 159 460,80