TY - JOUR
T1 - Characterization of Trypanosoma cruzi MutY DNA glycosylase ortholog and its role in oxidative stress response
AU - Kunrath-Lima, Marianna
AU - Repolês, Bruno Marçal
AU - Alves, Ceres Luciana
AU - Furtado, Carolina
AU - Rajão, Matheus Andrade
AU - Macedo, Andrea Mara
AU - Franco, Glória Regina
AU - Pena, Sérgio Danilo Junho
AU - Valenzuela, Lucía
AU - Wisnovsky, Simon
AU - Kelley, Shana O.
AU - Galanti, Norbel
AU - Cabrera, Gonzalo
AU - Machado, Carlos Renato
N1 - Funding Information:
This work was supported by FONDECYT (Chile) [grant number 1130113 ]; ERANET-LAC [grant number ELAC2014/HID-0328 ]; CNPq (Brazil) [grant number 444334/2014-9 ]; FAPEMIG (Brazil) [grant numbers APQ-00827-15 , APQ-02308-13 ].
Publisher Copyright:
© 2017 Elsevier B.V.
PY - 2017/11
Y1 - 2017/11
N2 - 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.
AB - 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.
KW - DNA repair
KW - MutY DNA glycosylase
KW - Oxidative stress
KW - Trypanosoma cruzi
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U2 - 10.1016/j.meegid.2017.09.030
DO - 10.1016/j.meegid.2017.09.030
M3 - Article
C2 - 28970112
AN - SCOPUS:85030833008
SN - 1567-1348
VL - 55
SP - 332
EP - 342
JO - Infection, Genetics and Evolution
JF - Infection, Genetics and Evolution
ER -
