Depletion of the thioredoxin homologue tryparedoxin impairs antioxidative defence in African trypanosomes

In trypanosomes, the thioredoxin-type protein TXN (tryparedoxin) is a multi-purpose oxidoreductase that is involved in the detoxification of hydroperoxides, the synthesis of DNA precursors and the replication of the kinetoplastid DNA. African trypanosomes possess two isoforms that are localized in the cytosol and in the mitochondrion of the parasites respectively. Here we report on the biological significance of the cTXN (cytosolic TXN) of Trypanosoma brucei for hydroperoxide detoxification. Depending on the growth phase, the concentration of the protein is 3-7-fold higher in the parasite form infecting mammals (50-100 mu M) than in the form hosted by the tsetse fly (7-34 mu M). Depletion of the mRNA in bloodstream trypanosomes by RNA interference revealed the indispensability of the protein. Proliferation and viability of cultured trypanosomes were impaired when TXN was lowered to 1 M for more than 48 h. Although the levels of glutathione, glutathionylspermidine and trypanothione were increased 2-3.5-fold, the sensitivity against mu exogenously generated H2O2 was significantly enhanced. The results prove the essential role of the cTXN and its pivotal function in the parasite defence against oxidative stress.