The heterologous expression of Escherichia coli MutT enzyme is involved in the protection against oxidative stress in Leishmania braziliensis
| Autor: | Antônio Vinícius de Assis, Silvane M. F. Murta, Edward Oliveira, Carlos Renato Machado, Ceres Luciana Alves, Ana Maria Murta Santi, Laila de Carvalho Andrade, Wesley Roger Rodrigues Ferreira |
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| Rok vydání: | 2020 |
| Předmět: |
Antimony
Microbiology (medical) Guanine DNA repair DNA damage RC955-962 030231 tropical medicine Antiprotozoal Agents MutT 8-oxoG medicine.disease_cause Microbiology Leishmania braziliensis Mice 03 medical and health sciences 0302 clinical medicine Arctic medicine. Tropical medicine Escherichia coli medicine Animals Humans oxidative stress Pyrophosphatases Leishmania infantum biology Superoxide Dismutase Escherichia coli Proteins DNA replication Hydrogen Peroxide Cell cycle biology.organism_classification Molecular biology QR1-502 Rats Oxidative Stress Original Article Rabbits Heterologous expression Oxidative stress |
| Zdroj: | Memórias do Instituto Oswaldo Cruz, Volume: 115, Article number: e190469, Published: 06 JUL 2020 Memórias do Instituto Oswaldo Cruz Memórias do Instituto Oswaldo Cruz., Vol 115 (2020) |
| ISSN: | 1678-8060 0074-0276 |
| DOI: | 10.1590/0074-02760190469 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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