%0 Unpublished work %T Single locus phosphoproteomics reveals phosphorylation of RPA-1 is required for generation of single-strand DNA following a break at a subtelomeric locus %+ Biologie moléculaire des Trypanosomes - Trypanosome Molecular Biology %+ Plateforme de Protéomique / Proteomics platform %+ Hub Bioinformatique et Biostatistique - Bioinformatics and Biostatistics HUB %+ Division of Biomedical and Life Sciences, Faculty of Health and Medicine %A Mclaughlin, Emilia %A Dujeancourt-Henry, Annick %A Chaze, Thibault %A Gianetto, Quentin Giai %A Matondo, Mariette %A Urbaniak, Michael, D %A Glover, Lucy %Z Work in the LG laboratory is has received financial support from the Institut Pasteur (G5 Junior group). EJM is part of the Pasteur - Paris University (PPU) International PhD Program. This project has received funding from the European Union's Horizon 2020 research and innovation programme under the Marie Sklodowska-Curie grant agreement No 665807 and from the Foundation Recherché Médicale grant number FDT202012010602. Funding for open access charge: Institut Pasteur G5 funding. %8 2023-05-10 %D 2023 %R 10.1101/2022.06.15.496243 %K Phosphoproteomics %K Trypanosoma brucei %K DNA damage %Z Life Sciences [q-bio]Preprints, Working Papers, ... %X Damage to the genetic material of the cell poses a universal threat to all forms of life. Central to the DNA damage response (DDR) is a phosphorylation signalling cascade that leads to the co-ordination of the cellular response to a DNA break. Identifying the proteins that are phosphorylated is crucial to understanding the mechanisms underlying this DDR. We have used SILAC-based quantitative phosphoproteomics to profile changes in phosphorylation site abundance following a single double strand break (DSB) at a chromosome internal locus and the subtelomeric bloodstream form expression site in Trypanosoma brucei . We report >6500 phosphorylation sites, including a core set of 211 DSB responsive phosphorylation sites. Along with phosphorylation of canonical DNA damage factors, we find that there is a striking distinction between the proteins phosphorylated in response to a chromosome internal DSB and one at the active BES and describe a single phosphorylation event on Replication factor A (RPA) 1 that is required for efficient resection at a bloodstream form expression site. %G English %Z EJM, MM, MU and LG designed the experiments. EJM, ADH, TC performed the experiments,EJM, QGG and MU performed the statistical analysis. EJM and LG wrote the manuscript, all authors edited the manuscript. We would also like to thank Sebastian Hutchinson for his guidance with the analysis of the VSG-sequencing %2 https://pasteur.hal.science/pasteur-04093539/document %2 https://pasteur.hal.science/pasteur-04093539/file/2022.06.15.496243.full.pdf %L pasteur-04093539 %U https://pasteur.hal.science/pasteur-04093539 %~ PASTEUR %~ CNRS %~ OPENAIRE %~ INC-CNRS %~ UNIV-PARIS %~ UNIVERSITE-PARIS %~ TEST2-HALCNRS %~ FRANCE-GENOMIQUE %~ BIOINFO_BIOSTAT_HUB