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The heterologous expression of Escherichia coli MutT enzyme is involved in the protection against oxidative stress in Leishmania braziliensis
Andrade, Laila de Carvalho; Santi, Ana Maria Murta; Departamento de Bioquímica e ImunologiaAlves, Ceres Luciana; Departamento de Bioquímica e ImunologiaFerreira, Wesley Roger Rodrigues; Departamento de Bioquímica e Imunologiade Assis, Antônio Vinícius; Oliveira, Edward; Departamento de Bioquímica e ImunologiaMachado, Carlos Renato; Murta, Silvane Maria Fonseca.
  • Andrade, Laila de Carvalho; Fundação Oswaldo Cruz-Fiocruz. Instituto René Rachou. Belo Horizonte. BR
  • Santi, Ana Maria Murta; Fundação Oswaldo Cruz-Fiocruz. Instituto René Rachou. Belo Horizonte. BR
  • Departamento de Bioquímica e ImunologiaAlves, Ceres Luciana; Universidade Federal de Minas Gerais. Instituto de Ciências Biológicas. Departamento de Bioquímica e ImunologiaAlves, Ceres Luciana. Belo Horizonte. BR
  • Departamento de Bioquímica e ImunologiaFerreira, Wesley Roger Rodrigues; Universidade Federal de Minas Gerais. Instituto de Ciências Biológicas. Departamento de Bioquímica e ImunologiaFerreira, Wesley Roger Rodrigues. Belo Horizonte. BR
  • Departamento de Bioquímica e Imunologiade Assis, Antônio Vinícius; Universidade Federal de Minas Gerais. Instituto de Ciências Biológicas. Departamento de Bioquímica e Imunologiade Assis, Antônio Vinícius. Belo Horizonte. BR
  • Oliveira, Edward; Fundação Oswaldo Cruz-Fiocruz. Instituto René Rachou. Belo Horizonte. BR
  • Departamento de Bioquímica e ImunologiaMachado, Carlos Renato; Universidade Federal de Minas Gerais. Instituto de Ciências Biológicas. Departamento de Bioquímica e ImunologiaMachado, Carlos Renato. Belo Horizonte. BR
  • Murta, Silvane Maria Fonseca; Fundação Oswaldo Cruz-Fiocruz. Instituto René Rachou. Belo Horizonte. BR
Mem. Inst. Oswaldo Cruz ; 115: e190469, 2020. graf
Article in English | LILACS, SES-SP | ID: biblio-1135243
ABSTRACT
BACKGROUND Oxidative stress is responsible for generating DNA lesions and the 8-oxoguanine (8-oxoG) is the most commonly lesion found in DNA damage. When this base is incorporated during DNA replication, it could generate double-strand DNA breaks and cellular death. MutT enzyme hydrolyzes the 8-oxoG from the nucleotide pool, preventing its incorporation during DNA replication. OBJECTIVES To investigate the importance of 8-oxoG in Leishmania infantum and L. braziliensis, in this study we analysed the impact of heterologous expression of Escherichia coli MutT (EcMutT) enzyme in drug-resistance phenotype and defense against oxidative stress. METHODS Comparative analysis of L. braziliensis and L. infantum H2O2 tolerance and cell cycle profile were performed. Lines of L. braziliensis and L. infantum expressing EcMutT were generated and evaluated using susceptibility tests to H2O2 and SbIII, cell cycle analysis, γH2A western blotting, and BrdU native detection assay. FINDINGS Comparative analysis of tolerance to oxidative stress generated by H2O2 showed that L. infantum is more tolerant to exogenous H2O2 than L. braziliensis. In addition, cell cycle analysis showed that L. infantum, after treatment with H2O2, remains in G1 phase, returning to its normal growth rate after 72 h. In contrast, after treatment with H2O2, L. braziliensis parasites continue to move to the next stages of the cell cycle. Expression of the E. coli MutT gene in L. braziliensis and L. infantum does not interfere in parasite growth or in susceptibility to SbIII. Interestingly, we observed that L. braziliensis EcMutT-expressing clones were more tolerant to H2O2 treatment, presented lower activation of γH2A, a biomarker of genotoxic stress, and lower replication stress than its parental non-transfected parasites. In contrast, the EcMutT is not involved in protection against oxidative stress generated by H2O2 in L. infantum. MAIN CONCLUSIONS Our results showed that 8-oxoG clearance in L. braziliensis is important to avoid misincorporation during DNA replication after oxidative stress generated by H2O2.
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Full text: Available Index: LILACS (Americas) Main subject: Pyrophosphatases / Superoxide Dismutase / Leishmania braziliensis / Leishmania infantum / Escherichia coli Proteins / Escherichia coli / Guanine / Antimony Limits: Animals / Humans Country/Region as subject: South America / Brazil Language: English Journal: Mem. Inst. Oswaldo Cruz Year: 2020 Type: Article Institution/Affiliation country: Fundação Oswaldo Cruz-Fiocruz/BR / Universidade Federal de Minas Gerais/BR

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Full text: Available Index: LILACS (Americas) Main subject: Pyrophosphatases / Superoxide Dismutase / Leishmania braziliensis / Leishmania infantum / Escherichia coli Proteins / Escherichia coli / Guanine / Antimony Limits: Animals / Humans Country/Region as subject: South America / Brazil Language: English Journal: Mem. Inst. Oswaldo Cruz Year: 2020 Type: Article Institution/Affiliation country: Fundação Oswaldo Cruz-Fiocruz/BR / Universidade Federal de Minas Gerais/BR