Contributors: Uniwersytet Wroclawski = University of Wroclaw; Medical University of Lublin; Institut Gustave Roussy (IGR); Institut de Génétique et Développement de Rennes (IGDR); Université de Rennes (UR)-Centre National de la Recherche Scientifique (CNRS)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ); Intégrité du génome et cancers (IGC); École Pratique des Hautes Études (EPHE); Université Paris Sciences et Lettres (PSL)-Université Paris Sciences et Lettres (PSL)-Institut Gustave Roussy (IGR)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS); This work was financially supported by a grant from the National Science Centre (NCN), Poland, no. 2016/21/D/NZ1/00285 to E. B. and an ANR JCJC grant to G. R. (ANR-12-JSV8-0003). E. B. acknowledges the French Government and the Embassy of France in Poland, and the Foundation for Polish Sciences (FNP). N. L. received a French Government Scholarship (BGF) to perform a joint PhD project, a STER NAWA scholarship and a mobility grant from Région Bretagne, France. Fig. 1, Fig. 2, Fig. 4, and Graphical abstract were created with BioRender.com under a granted license to E. B.; ANR-12-JSV8-0003,NucUb,Ubiquitylation nucléaire(2012)
نبذة مختصرة : International audience ; In response to genotoxic stress, cells evolved with a complex signaling network referred to as the DNA damage response (DDR). It is now well established that the DDR depends upon various post-translational modifications; among them, ubiquitylation plays a key regulatory role. Here, we profiled ubiquitylation in response to the DNA alkylating agent methyl methanesulfonate (MMS) in the budding yeast Saccharomyces cerevisiae using quantitative proteomics. To discover new proteins ubiquitylated upon DNA replication stress, we used stable isotope labeling by amino acids in cell culture (SILAC) followed by an enrichment of ubiquitylated peptides and liquid chromatography-tandem mass spectrometry (LC-MS/MS). In total, we identified 1853 ubiquitylated proteins, including 473 proteins that appeared upregulated more than 2-fold in response to MMS treatment. This enabled us to localize 519 ubiquitylation sites potentially regulated upon MMS in 435 proteins. We demonstrated that the overexpression of some of these proteins renders the cells sensitive to MMS. We also assayed the abundance change upon MMS treatment of a selection of yeast nuclear proteins. Several of them were differentially regulated upon MMS treatment. These findings corroborate the important role of ubiquitin-proteasome-mediated degradation in regulating the DDR.
Processing Request
No Comments.