Mem Inst Oswaldo Cruz, Rio de Janeiro, VOLUME 115 | JUNE 2020
Original Article

Role of Trypanosoma cruzi nucleoside diphosphate kinase 1 in DNA-damage responses

Chantal Reigada1,2, Melisa Sayé1,2, Fabio Di Girolamo1,2, Edward A Valera-Vera1,2, Claudio A Pereira1,2, Mariana R Miranda1,2,+

1Universidad de Buenos Aires, Facultad de Medicina, Instituto de Investigaciones Médicas A Lanari, Buenos Aires, Argentina
2Consejo Nacional de Investigaciones Científicas y Técnicas, Universidad de Buenos Aires, Instituto de Investigaciones Médicas, Laboratorio de Parasitología Molecular, Buenos Aires, Argentina

DOI: 10.1590/0074-02760200019
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ABSTRACT

BACKGROUND NME23/NDPKs are well conserved proteins found in all living organisms. In addition to being nucleoside diphosphate kinases (NDPK), they are multifunctional enzymes involved in different processes such as DNA stability, gene regulation and DNA repair among others. TcNDPK1 is the canonical NDPK isoform present in Trypanosoma cruzi, which has nuclease activity and DNA-binding properties in vitro.
OBJECTIVES In the present study we explored the role of TcNDPK1 in DNA damage responses.
METHODS TcNDPK1 was expressed in mutant bacteria and yeasts and over-expressed in epimastigotes. Mutation frequencies, tolerance to genotoxic agents and activity of DNA repair enzymes were evaluated.
FINDINGS Bacteria decreased about 15-folds the spontaneous mutation rate and yeasts were more resistant to hydrogen peroxide and to UV radiation than controls. Parasites overexpressing TcNDPK1 were able to withstand genotoxic stresses caused by peroxide, phleomycin and hidroxyurea. They also presented less genomic damage and augmented levels of poly(ADP) ribose and poly(ADP)ribose polymerase, an enzyme involved in DNA repair.
MAIN CONCLUSION These results strongly suggest a novel function for TcNDPK1; its involvement in the maintenance of parasite’s genome integrity.

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Financial support: This work was supported by Consejo Nacional de Investigaciones Científicas y Técnicas and Agencia Nacional de Promoción Científica y Tecnológica (ANPCyT) (grants: FONCYT PICT 2013-2218, 2015-0539, 2018-1801 and 2018-1871). The research leading to these results has, in part, received funding from UK Research and Innovation via the Global Challenges Research Fund under grant agreement ‘A Global Network for Neglected Tropical Diseases’ (grant number MR/P027989/1). CONICET for salaries of researchers, fellowships and technicians: CAP, MRM, CR, FDG and EV-V. MS is a PDRA from A Global Network for Neglected Tropical Diseases.
CR and MS contributed equally to this work.
+ Corresponding author: mmiranda@conicet.gov / mirandamariana@gmail.com
ORCID http://orcid.org/0000-0002-0692-5675
Received 15 January 2020
Accepted 02 June 2020

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