The in vivo and in vitro roles of Trypanosoma cruzi Rad51 in the repair of DNA double strand breaks and oxidative lesions.

• Published in: PLOS Neglected Tropical Diseases
• Main Authors: Danielle Gomes Passos Silva, Selma da Silva Santos, Sheila C Nardelli, Isabela Cecília Mendes, Anna Cláudia Guimarães Freire, Bruno Marçal Repolês, Bruno Carvalho Resende, Héllida Marina Costa-Silva, Verônica Santana da Silva, Karla Andrade de Oliveira, Camila Franco Batista Oliveira, Liza Figueiredo Felicori Vilela, Ronaldo Alves Pinto Nagem, Glória Regina Franco, Andrea Mara Macedo, Sergio Danilo Junho Pena, Erich Birelli Tahara, Policarpo Ademar Sales Junior, Douglas Souza Moreira, Santuza Maria Ribeiro Teixeira, Richard McCulloch, Stela Virgilio, Luiz Ricardo Orsini Tosi, Sergio Schenkman, Luciana Oliveira Andrade, Silvane Maria Fonseca Murta, Carlos Renato Machado
• Format: Article in Journal/Newspaper
• Language: English
• Published: Public Library of Science (PLoS) 2018
• Subjects: Arctic medicine. Tropical medicine; RC955-962; Public aspects of medicine; RA1-1270; Arctic
• Online Access: https://doi.org/10.1371/journal.pntd.0006875; https://doaj.org/article/36c29e39f2654429899460add329f872

In Trypanosoma cruzi, the etiologic agent of Chagas disease, Rad51 (TcRad51) is a central enzyme for homologous recombination. Here we describe the different roles of TcRad51 in DNA repair. Epimastigotes of T. cruzi overexpressing TcRAD51 presented abundant TcRad51-labeled foci before gamma irradiation treatment, and a faster growth recovery when compared to single-knockout epimastigotes for RAD51. Overexpression of RAD51 also promoted increased resistance against hydrogen peroxide treatment, while the single-knockout epimastigotes for RAD51 exhibited increased sensitivity to this oxidant agent, which indicates a role for this gene in the repair of DNA oxidative lesions. In contrast, TcRad51 was not involved in the repair of crosslink lesions promoted by UV light and cisplatin treatment. Also, RAD51 single-knockout epimastigotes showed a similar growth rate to that exhibited by wild-type ones after treatment with hydroxyurea, but an increased sensitivity to methyl methane sulfonate. Besides its role in epimastigotes, TcRad51 is also important during mammalian infection, as shown by increased detection of T. cruzi cells overexpressing RAD51, and decreased detection of single-knockout cells for RAD51, in both fibroblasts and macrophages infected with amastigotes. Besides that, RAD51-overexpressing parasites infecting mice also presented increased infectivity and higher resistance against benznidazole. We thus show that TcRad51 is involved in the repair of DNA double strands breaks and oxidative lesions in two different T. cruzi developmental stages, possibly playing an important role in the infectivity of this parasite.