Girasol, Mark John and Krasilnikova, Marija and Marques, Catarina A. and Damasceno, Jeziel D. and Lapsley, Craig and Lemgruber, Leandro and Burchmore, Richard and Beraldi, Dario and Carruthers, Ross and Briggs, Emma M. and McCulloch, Richard (2023) RAD51-mediated R-loop formation acts to repair transcription associated DNA breaks driving antigenic variation in Trypanosoma brucei. Proceedings of the National Academy of Sciences of the United States of America, 120 (48): e230930612. ISSN 0027-8424

Abstract

RNA–DNA hybrids are epigenetic features of all genomes that intersect with many processes, including transcription, telomere homeostasis, and centromere function. Increasing evidence suggests that RNA–DNA hybrids can provide two conflicting roles in the maintenance and transmission of genomes: They can be the triggers of DNA damage, leading to genome change, or can aid the DNA repair processes needed to respond to DNA lesions. Evasion of host immunity by African trypanosomes, such as Trypanosoma brucei, relies on targeted recombination of silent Variant Surface Glycoprotein (VSG) genes into a specialized telomeric locus that directs transcription of just one VSG from thousands. How such VSG recombination is targeted and initiated is unclear. Here, we show that a key enzyme of T. brucei homologous recombination, RAD51, interacts with RNA–DNA hybrids. In addition, we show that RNA–DNA hybrids display a genome-wide colocalization with DNA breaks and that this relationship is impaired by mutation of RAD51. Finally, we show that RAD51 acts to repair highly abundant, localised DNA breaks at the single transcribed VSG and that mutation of RAD51 alters RNA–DNA hybrid abundance at 70 bp repeats both around the transcribed VSG and across the silent VSG archive. This work reveals a widespread, generalised role for RNA–DNA hybrids in directing RAD51 activity during recombination and uncovers a specialised application of this interplay during targeted DNA break repair needed for the critical T. brucei immune evasion reaction of antigenic variation.

People
Girasol, Mark John
Author

Girasol, Mark John and Krasilnikova, Marija and Marques, Catarina A. and Damasceno, Jeziel D. and Lapsley, Craig and Lemgruber, Leandro and Burchmore, Richard and Beraldi, Dario and Carruthers, Ross and Briggs, Emma M. and McCulloch, Richard (2023) RAD51-mediated R-loop formation acts to repair transcription associated DNA breaks driving antigenic variation in Trypanosoma brucei. Proceedings of the National Academy of Sciences of the United States of America, 120 (48): e230930612. ISSN 0027-8424

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Krasilnikova, Marija
Author

Girasol, Mark John and Krasilnikova, Marija and Marques, Catarina A. and Damasceno, Jeziel D. and Lapsley, Craig and Lemgruber, Leandro and Burchmore, Richard and Beraldi, Dario and Carruthers, Ross and Briggs, Emma M. and McCulloch, Richard (2023) RAD51-mediated R-loop formation acts to repair transcription associated DNA breaks driving antigenic variation in Trypanosoma brucei. Proceedings of the National Academy of Sciences of the United States of America, 120 (48): e230930612. ISSN 0027-8424

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Marques, Catarina A.
Author

Girasol, Mark John and Krasilnikova, Marija and Marques, Catarina A. and Damasceno, Jeziel D. and Lapsley, Craig and Lemgruber, Leandro and Burchmore, Richard and Beraldi, Dario and Carruthers, Ross and Briggs, Emma M. and McCulloch, Richard (2023) RAD51-mediated R-loop formation acts to repair transcription associated DNA breaks driving antigenic variation in Trypanosoma brucei. Proceedings of the National Academy of Sciences of the United States of America, 120 (48): e230930612. ISSN 0027-8424

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Damasceno, Jeziel D.
Author

Girasol, Mark John and Krasilnikova, Marija and Marques, Catarina A. and Damasceno, Jeziel D. and Lapsley, Craig and Lemgruber, Leandro and Burchmore, Richard and Beraldi, Dario and Carruthers, Ross and Briggs, Emma M. and McCulloch, Richard (2023) RAD51-mediated R-loop formation acts to repair transcription associated DNA breaks driving antigenic variation in Trypanosoma brucei. Proceedings of the National Academy of Sciences of the United States of America, 120 (48): e230930612. ISSN 0027-8424

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Lapsley, Craig
Author

Girasol, Mark John and Krasilnikova, Marija and Marques, Catarina A. and Damasceno, Jeziel D. and Lapsley, Craig and Lemgruber, Leandro and Burchmore, Richard and Beraldi, Dario and Carruthers, Ross and Briggs, Emma M. and McCulloch, Richard (2023) RAD51-mediated R-loop formation acts to repair transcription associated DNA breaks driving antigenic variation in Trypanosoma brucei. Proceedings of the National Academy of Sciences of the United States of America, 120 (48): e230930612. ISSN 0027-8424

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Lemgruber, Leandro
Author

Girasol, Mark John and Krasilnikova, Marija and Marques, Catarina A. and Damasceno, Jeziel D. and Lapsley, Craig and Lemgruber, Leandro and Burchmore, Richard and Beraldi, Dario and Carruthers, Ross and Briggs, Emma M. and McCulloch, Richard (2023) RAD51-mediated R-loop formation acts to repair transcription associated DNA breaks driving antigenic variation in Trypanosoma brucei. Proceedings of the National Academy of Sciences of the United States of America, 120 (48): e230930612. ISSN 0027-8424

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Burchmore, Richard
Author

Girasol, Mark John and Krasilnikova, Marija and Marques, Catarina A. and Damasceno, Jeziel D. and Lapsley, Craig and Lemgruber, Leandro and Burchmore, Richard and Beraldi, Dario and Carruthers, Ross and Briggs, Emma M. and McCulloch, Richard (2023) RAD51-mediated R-loop formation acts to repair transcription associated DNA breaks driving antigenic variation in Trypanosoma brucei. Proceedings of the National Academy of Sciences of the United States of America, 120 (48): e230930612. ISSN 0027-8424

See full publications list
Beraldi, Dario
Author

Girasol, Mark John and Krasilnikova, Marija and Marques, Catarina A. and Damasceno, Jeziel D. and Lapsley, Craig and Lemgruber, Leandro and Burchmore, Richard and Beraldi, Dario and Carruthers, Ross and Briggs, Emma M. and McCulloch, Richard (2023) RAD51-mediated R-loop formation acts to repair transcription associated DNA breaks driving antigenic variation in Trypanosoma brucei. Proceedings of the National Academy of Sciences of the United States of America, 120 (48): e230930612. ISSN 0027-8424

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Carruthers, Ross
Author

Briceño Guel, Diana Nohemi and Laverick, Nicola and MacLaren, Linda and MacLeod, Nicholas and Glegg, Martin and Lamb, Gillian and Houston, Peter and Carruthers, Ross and Grocutt, Laura and Valentine, Ronan (2024) Adaptive radiotherapy for muscle invasive bladder cancer: a retrospective audit of two bladder filling protocols. Radiation Oncology, 19 (1): 92. ISSN 1748-717X

Girasol, Mark John and Krasilnikova, Marija and Marques, Catarina A. and Damasceno, Jeziel D. and Lapsley, Craig and Lemgruber, Leandro and Burchmore, Richard and Beraldi, Dario and Carruthers, Ross and Briggs, Emma M. and McCulloch, Richard (2023) RAD51-mediated R-loop formation acts to repair transcription associated DNA breaks driving antigenic variation in Trypanosoma brucei. Proceedings of the National Academy of Sciences of the United States of America, 120 (48): e230930612. ISSN 0027-8424

See full publications list
Briggs, Emma M.
Author

Girasol, Mark John and Krasilnikova, Marija and Marques, Catarina A. and Damasceno, Jeziel D. and Lapsley, Craig and Lemgruber, Leandro and Burchmore, Richard and Beraldi, Dario and Carruthers, Ross and Briggs, Emma M. and McCulloch, Richard (2023) RAD51-mediated R-loop formation acts to repair transcription associated DNA breaks driving antigenic variation in Trypanosoma brucei. Proceedings of the National Academy of Sciences of the United States of America, 120 (48): e230930612. ISSN 0027-8424

See full publications list
McCulloch, Richard
Author

Girasol, Mark John and Krasilnikova, Marija and Marques, Catarina A. and Damasceno, Jeziel D. and Lapsley, Craig and Lemgruber, Leandro and Burchmore, Richard and Beraldi, Dario and Carruthers, Ross and Briggs, Emma M. and McCulloch, Richard (2023) RAD51-mediated R-loop formation acts to repair transcription associated DNA breaks driving antigenic variation in Trypanosoma brucei. Proceedings of the National Academy of Sciences of the United States of America, 120 (48): e230930612. ISSN 0027-8424

See full publications list
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Information
Title:
RAD51-mediated R-loop formation acts to repair transcription associated DNA breaks driving antigenic variation in Trypanosoma brucei
Creators:
Girasol, Mark John and Krasilnikova, Marija and Marques, Catarina A. and Damasceno, Jeziel D. and Lapsley, Craig and Lemgruber, Leandro and Burchmore, Richard and Beraldi, Dario and Carruthers, Ross and Briggs, Emma M. and McCulloch, Richard
Identification Number:
10.1073/pnas.2309306120
Date:
28 November 2023
Divisions:
College of Medical Veterinary and Life Sciences
College of Medical Veterinary and Life Sciences > School of Cancer Sciences
Publisher:
National Academy of Sciences
Additional Information:
This work was funded by the Philippine Council for Health Research and Development (M.J.G.), a Wellcome Trust Investigator Award (224501/Z/21/Z, R.M.), a Henry Wellcome fellowship (218648/Z/19/Z, E.M.B.), an Institutional Strategic Support Fund award to the University of Glasgow (204820/Z/16/Z), and by the BBSRC (BB/N016165/1 and BB/R017166/1, BB/W001101/1, C.A.M. and R.M.). The Wellcome Centre for Integrative Parasitology is funded by a Wellcome Trust Strategic Award (104111/Z/14/Z/A).
Library
URI:
https://pub.demo35.eprints-hosting.org/id/eprint/453
Item Type:
Article
Depositing User:
Unnamed user with email ejo1f20@soton.ac.uk
Date Deposited:
11 Jun 2025 16:38
Revision:
45
Last Modified:
12 Jun 2025 09:20
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