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Single locus phosphoproteomics reveals phosphorylation of RPA-1 is required for generation of single-strand DNA following a break at a subtelomeric locus

Emilia McLaughlin, Annick Dujeancourt-Henry, Thibault Chaze, Quentin Giai Gianetto, Mariette Matondo, View ORCID ProfileMichael D Urbaniak, Lucy Glover
doi: https://doi.org/10.1101/2022.06.15.496243
Emilia McLaughlin
1Institut Pasteur, Université Paris Cité, Trypanosome Molecular Biology, Department of Parasites and Insect Vectors, F-75015, Paris, France
2Sorbonne Université, Collège doctoral, F-75005 Paris, France
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Annick Dujeancourt-Henry
1Institut Pasteur, Université Paris Cité, Trypanosome Molecular Biology, Department of Parasites and Insect Vectors, F-75015, Paris, France
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Thibault Chaze
3Institut Pasteur, Université Paris Cité, Proteomics Platform, Mass Spectrometry for Biology Unit, Centre National de la Recherche Scientifique, UAR 2024, 75015 Paris, France
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Quentin Giai Gianetto
3Institut Pasteur, Université Paris Cité, Proteomics Platform, Mass Spectrometry for Biology Unit, Centre National de la Recherche Scientifique, UAR 2024, 75015 Paris, France
4Institut Pasteur, Université Paris Cité, Bioinformatics and Biostatistics HUB, 75015 Paris, France
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Mariette Matondo
3Institut Pasteur, Université Paris Cité, Proteomics Platform, Mass Spectrometry for Biology Unit, Centre National de la Recherche Scientifique, UAR 2024, 75015 Paris, France
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Michael D Urbaniak
5Division of Biomedical and Life Sciences, Faculty of Health and Medicine, Lancaster University, Lancaster, UK
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  • ORCID record for Michael D Urbaniak
Lucy Glover
1Institut Pasteur, Université Paris Cité, Trypanosome Molecular Biology, Department of Parasites and Insect Vectors, F-75015, Paris, France
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  • For correspondence: lucy.glover@pasteur.fr
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Abstract

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.

Competing Interest Statement

The authors have declared no competing interest.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-ND 4.0 International license.
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Posted June 15, 2022.
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Single locus phosphoproteomics reveals phosphorylation of RPA-1 is required for generation of single-strand DNA following a break at a subtelomeric locus
Emilia McLaughlin, Annick Dujeancourt-Henry, Thibault Chaze, Quentin Giai Gianetto, Mariette Matondo, Michael D Urbaniak, Lucy Glover
bioRxiv 2022.06.15.496243; doi: https://doi.org/10.1101/2022.06.15.496243
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Single locus phosphoproteomics reveals phosphorylation of RPA-1 is required for generation of single-strand DNA following a break at a subtelomeric locus
Emilia McLaughlin, Annick Dujeancourt-Henry, Thibault Chaze, Quentin Giai Gianetto, Mariette Matondo, Michael D Urbaniak, Lucy Glover
bioRxiv 2022.06.15.496243; doi: https://doi.org/10.1101/2022.06.15.496243

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