Characterization of Trypanosoma cruzi MutY DNA glycosylase ortholog and its role in oxidative stress response

Marianna Kunrath-Lima, Bruno Marçal Repolês, Ceres Luciana Alves, Carolina Furtado, Matheus Andrade Rajão, Andrea Mara Macedo, Glória Regina Franco, Sérgio Danilo Junho Pena, Lucía Valenzuela, Simon Wisnovsky, Shana O. Kelley, Norbel Galanti, Gonzalo Cabrera, Carlos Renato Machado*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

4 Scopus citations

Abstract

Trypanosoma cruzi is a protozoan parasite and the causative agent of Chagas disease. Like most living organisms, it is susceptible to oxidative stress, and must adapt to distinct environments. Hence, DNA repair is essential for its survival and the persistence of infection. Therefore, we studied whether T. cruzi has a homolog counterpart of the MutY enzyme (TcMYH), important in the DNA Base Excision Repair (BER) mechanism. Analysis of T. cruzi genome database showed that this parasite has a putative MutY DNA glycosylase sequence. We performed heterologous complementation assays using this genomic sequence. TcMYH complemented the Escherichia coli MutY- strain, reducing the mutation rate to a level similar to wild type. In in vitro assays, TcMYH was able to remove an adenine that was opposite to 8-oxoguanine. We have also constructed a T. cruzi lineage that overexpresses MYH. Although in standard conditions this lineage has similar growth to control cells, the overexpressor is more sensitive to hydrogen peroxide and glucose oxidase than the control, probably due to accumulation of AP sites in its DNA. Localization experiments with GFP-fused TcMYH showed this enzyme is present in both nucleus and mitochondrion. QPCR and MtOX results reinforce the presence and function of TcMYH in these two organelles. Our data suggest T. cruzi has a functional MYH DNA glycosylase, which participates in nuclear and mitochondrial DNA Base Excision Repair.

Original languageEnglish (US)
Pages (from-to)332-342
Number of pages11
JournalInfection, Genetics and Evolution
Volume55
DOIs
StatePublished - Nov 2017
Externally publishedYes

Keywords

  • DNA repair
  • MutY DNA glycosylase
  • Oxidative stress
  • Trypanosoma cruzi

ASJC Scopus subject areas

  • Microbiology
  • Ecology, Evolution, Behavior and Systematics
  • Molecular Biology
  • Genetics
  • Microbiology (medical)
  • Infectious Diseases

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