RAD51-mediated R-loop formation acts to repair transcription-associated DNA breaks driving antigenic variation in Trypanosoma brucei .

Publisher: National Academy of Sciences Country of Publication: United States NLM ID: 7505876 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1091-6490 (Electronic) Linking ISSN: 00278424 NLM ISO Abbreviation: Proc Natl Acad Sci U S A Subsets: MEDLINE

Original Publication: Washington, DC : National Academy of Sciences

Trypanosoma brucei brucei*; R-Loop Structures ; Antigenic Variation/genetics ; DNA Breaks ; DNA ; RNA ; Variant Surface Glycoproteins, Trypanosoma/genetics

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.
Competing Interests: Competing interests statement:The authors declare no competing interest.

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United Kingdom WT_ Wellcome Trust

Keywords: DNA repair; RNA–DNA hybrid; antigenic variation; recombination; trypanosome

9007-49-2 (DNA)
63231-63-0 (RNA)
0 (Variant Surface Glycoproteins, Trypanosoma)

Date Created: 20231121 Date Completed: 20231127 Latest Revision: 20231205

20231215

PMC10691351

10.1073/pnas.2309306120

37988471