Discussion

Protozoan parasite Trypanosoma brucei (Euglenozoa, Kinetoplastea, Trypanosomatida) is the causative agent of sleeping sickness in human and disease Nagano in animals. Better knowledge of this organism is the key for successful fight against caused diseases. Our research focuses on mitochondrial FtsH protease, which was not being closely studied among trypanosomatids yet.  FtsH protease is a representative of mitochondrial AAA proteases associated with various cell activities. In genome of T. brucei has been identified up to six homologs of FtsH subunits unlike S. cerevisiae and human which each possess only three different subunits. Subunits can form homo- or hetero- hexameric structures oriented into the mitochondrial matrix or intermembrane space, depending on transmembrane sequences on their N terminus. We have prepared cell lines with inducible RNAi of each subunit, and also V5 tagged cell lines. Influence of gene silencing on growth phenotype and activity of oxidative phosphorylation has been examined. We have also performed in silico analysis for the presence of transmembrane domains of these six genes, predicting their orientation.