Increased expression of iron-containing superoxide dismutase-A (TcFeSOD-A) enzyme in Trypanosoma cruzi population with in vitro-induced resistance to benznidazole.
Superoxide dismutase (SOD) removes excess superoxide radicals via dismutation to oxygen and hydrogen peroxide. In this work, we have characterized TcFeSOD-A gene from 25 Trypanosoma cruzi populations and clones susceptible, naturally resistant or with in vitro-induced (17 LER) or in vivo-selected resistance to benznidazole (BZR). In the 17 LER T. cruzi population, the levels of TcFeSOD-A mRNA were at least 3-fold higher than its drug-susceptible counterpart 17 WTS. The levels of TcFeSOD-A mRNA were similar among the other T. cruzi populations and clones regardless of the drug-resistance phenotype. We determined whether the increase in mRNA levels was due to gene amplification using Southern blot analysis of the T. cruzi populations and clones. We found that the number of TcFeSOD-A gene copies was similar for all samples tested, except for 17 LER that presented twice as many copies. The chromosomal location of the TcFeSOD-A gene and polymorphisms detected in nucleotide and amino acid sequences of TcFeSOD-A were associated with the zymodeme of the T. cruzi strain but not with drug-resistance phenotype. We observed a 23 kDa TcFeSOD-A polypeptide in all analysed T. cruzi strains. The level of this polypeptide was increased only in the 17 LER population. Specific enzyme activity analysis of TcFeSOD in the T. cruzi samples revealed a correlation between expression and activity. Our findings show an increased expression of the TcFeSOD-A enzyme in the T. cruzi population with in vitro-induced resistance to benznidazole.
Nogueira
Fernanda B
FB
Laboratório de Parasitologia Celular e Molecular, Centro de Pesquisas René Rachou, FIOCRUZ, Av Augusto de Lima 1715, 30190-002 Belo Horizonte, MG, Brazil.
Krieger
Marco A
MA
Nirdé
Philippe
P
Goldenberg
Samuel
S
Romanha
Alvaro J
AJ
Murta
Silvane M F
SM
D000595:Amino Acid Sequence; D000818:Animals; D002874:Chromosome Mapping; D004351:Drug Resistance; D018628:Gene Dosage; D006801:Humans; D008969:Molecular Sequence Data; D009593:Nitroimidazoles / Q000494:pharmacology*; D021261:Parasitic Sensitivity Tests; D017422:Sequence Analysis, DNA; D013482:Superoxide Dismutase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D014344:Trypanocidal Agents / Q000494:pharmacology*; D014349:Trypanosoma cruzi / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D015854:Up-Regulation
D009593:Nitroimidazoles; D014344:Trypanocidal Agents; D013482:Superoxide Dismutase; C009999:benzonidazole