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

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 hydrogen 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.

X-ray diffraction and in vivo studies reveal the quinary structure of Trypanosoma cruzi nucleoside diphosphate kinase 1: a novel helical oligomer structure

“…Previous reports have shown that TcNDPK1 has nuclease activity in vitro (Miranda et al, 2008). More recently, Reigada et al (2020) demonstrated the participation of TcNDPK1 in DNA-damage responses. Like most eukaryotic NDPKs, TcNDPK1 forms hexamers (Ulloa et al, 1995;Souza et al, 2011).…”

“…Other NDPKs from pathogens such as parasites are involved in proliferation (Castellanos-Gonzalez et al, 2019). They act as virulence factors (Kolli et al, 2008) and participate in responses to DNA damage (Reigada et al, 2020). Despite many studies, the multiple functions of these enzymes are not fully understood.…”

Revisiting trypanosomatid nucleoside diphosphate kinases

“…As a result, a wide variety of unrelated processes from prokaryotes and eukaryotes have been reported in which canonical enzymes play a role. These processes include the following: (a) signal transduction, 27 , 28 (b) virulence and host-pathogen interactions, 29 , 30 , 31 , 32 (c) apoptosis, 33 (d) metastasis, 9 (e) gene regulation 34 and (f) genome integrity, 35 , 36 , 37 among others. 38 , 39 , 40 One intriguing role that deserves special attention, is the case of human NME2, first discovered as Puf, which acts as a transcription factor regulating c-myc expression through binding to a G-quadruplex DNA secondary structure within the promoter region.…”

“…In this regard, nuclease activity was confirmed for TcNDPK1 17 and, moreover, in a recent study, it was demonstrated that TcNDPK1 is involved in T. cruzi genome integrity maintenance, exerting a protective role against genotoxic agents probably through the up-regulation of enzymes involved in DNA repair. 35 Therefore, the benznidazole resistant phenotype could be associated with this TcNDPK1 function.…”