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RAD50 promotes DNA repair by homologous recombination and restrains antigenic variation in African trypanosomes

View ORCID ProfileAnn-Kathrin Mehnert, Marco Prorocic, Annick Dujeancourt-Henry, Sebastian Hutchinson, View ORCID ProfileRichard McCulloch, View ORCID ProfileLucy Glover
doi: https://doi.org/10.1101/2020.03.17.994905
Ann-Kathrin Mehnert
1Trypanosome Molecular Biology, Department of Parasites and Insect Vectors, Institut Pasteur, 25-28 Rue du Docteur Roux 75015, Paris, France
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  • ORCID record for Ann-Kathrin Mehnert
Marco Prorocic
2Wellcome Center for Integrative Parasitology, Sir Graeme Davis Building, 120 University Place, Glasgow, G12 8TA, UK
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Annick Dujeancourt-Henry
1Trypanosome Molecular Biology, Department of Parasites and Insect Vectors, Institut Pasteur, 25-28 Rue du Docteur Roux 75015, Paris, France
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Sebastian Hutchinson
3Trypanosome Cell Biology, Department of Parasites and Insect Vectors, Institut Pasteur, 25-28 Rue du Docteur Roux 75015, Paris, France
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Richard McCulloch
2Wellcome Center for Integrative Parasitology, Sir Graeme Davis Building, 120 University Place, Glasgow, G12 8TA, UK
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Lucy Glover
1Trypanosome Molecular Biology, Department of Parasites and Insect Vectors, Institut Pasteur, 25-28 Rue du Docteur Roux 75015, Paris, France
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  • For correspondence: lucy.glover@pasteur.fr
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ABSTRACT

Homologous recombination dominates as the major form of DNA repair in Trypanosoma brucei, and is especially important for recombination of the subtelomeric variant surface glycoprotein during antigenic variation. RAD50, a component of the MRN complex (MRE11, RAD50, NBS1), is central to homologous recombination through facilitating resection and governing the DNA damage response. The function of RAD50 in trypanosomes is untested. Here we report that RAD50 is required for RAD51-dependent homologous recombination, phosphorylation of histone H2A and controlled resection following a DNA double strand break (DSB). Perhaps surprisingly, DSB resection in the rad50 nulls was not impaired and appeared to peak earlier than in the parental strains. Finally, we show that RAD50 suppresses DNA repair using donors with short stretches of homology at a subtelomeric locus, with null strains producing a greater diversity of expressed VSG variants following DSB repair. We conclude that RAD50 promotes stringent homologous recombination at subtelomeric loci and restrains antigenic variation.

Footnotes

  • https://github.com/LGloverTMB/DNA-repair-mutant-VSG-seq

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission.
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RAD50 promotes DNA repair by homologous recombination and restrains antigenic variation in African trypanosomes
Ann-Kathrin Mehnert, Marco Prorocic, Annick Dujeancourt-Henry, Sebastian Hutchinson, Richard McCulloch, Lucy Glover
bioRxiv 2020.03.17.994905; doi: https://doi.org/10.1101/2020.03.17.994905
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RAD50 promotes DNA repair by homologous recombination and restrains antigenic variation in African trypanosomes
Ann-Kathrin Mehnert, Marco Prorocic, Annick Dujeancourt-Henry, Sebastian Hutchinson, Richard McCulloch, Lucy Glover
bioRxiv 2020.03.17.994905; doi: https://doi.org/10.1101/2020.03.17.994905

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