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Study of mitochondrial processing peptidases in procyclic \kur{Trypanosoma brucei}
POLIAK, Pavel
Aim of this work was to find out how mitochondrial processing peptidases are working in the mitochondrion of Trypanosoma brucei. I have shown by RNA interference that mitochondrial processing peptidase (MPP) and mitochondrial intermediate peptidase (MIP) are essential for procyclic stages. Moreover, processing of human frataxin in T. brucei has a similar pattern as in human cells.
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Functional analysis of Ssc1 and Iba57 proteins in \kur{Trypanosoma brucei}
SKALICKÝ, Tomáš
Aim of this thesis was to shed light on the function(s) of Iba57 and Ssc1 proteins in both life cycle stages of T. brucei using RNA interference. Depletion of Ssc1 resulted in severe grow phenotype, decrease in activities of iron-sulphur cluster-containing enzyme aconitase but no increase in oxidative stress sensitivity or accumulation of ROS in mitochondrion. Down regulation of Iba57, specialized maturation factor of aconitase and homoaconitase, lead to depletion of aconitase, destabilization of Isa1 and increased sensitivity to oxidative stress and accumulation of ROS in both stages.
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Characterization of two members from the multigenic family of one-domain Kunitz-inhibitors from the tick \kur{Ixodes ricinus}
SINGEROVÁ, Barbora
Two new genes encoding proteins Monolaris 1 and Monolaris 2 were isolated from tick Ixodes ricinus. Both cDNA fragments code for 94 aminoacid residues long protein with molecular mass 8,1kDa (Monolaris 1) and 8,3kDa (Monolaris 2). The function of Monolaris 1 was tested by using RNA interference in adult females of Ixodes ricinus that were subsequently fed on guinea-pigs. Body mass, egg mass and mortality were measured to evaluate the effect of gene silencing. Recombinant protein Monolaris 1 was prepared in bacterial expression system and antibodies against this protein were raised by immunization of a rabbit. Antibodies reacted with approximately 190kDa big protein in salivary gland, ovary and gut whereas monomer of Monolaris 1 was not detected in tick saliva, salivary glands and other tissues.
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Functional analysis of the YCF 45 gene in procyclic \kur{Trypanosoma brucei}
TÝČ, Jiří
In this study was shown by RNA interference that YCF 45 gene of putative chloroplast origin is not essential for surviving of procyclic Trypanosoma brucei. Three different construct bearing HA-tagged YCF 45 protein that differs on their N-terminus were generated to reveal the localization of the protein in the cell. Spliced leader acceptor site of YCF 45 mRNA was identified and in comparison to annotated data new start codon was proposed. Fylogeny tree was made to shed some light on the origin of this gene.
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The characterization of adenosine signal pathway in \kur{Drosophila} imaginal disc cells
TICHÝ, Vlastimil
The aim of this work was to characterise the influence of adenosine on imaginal disc cell line Cl8+ of Drosophila. I prepared stable cell lines with the overexpression or RNA interference of genes coding adenosine receptor AdoR (CG9753) and adenosine transporter DmENT2 (CG11045) in D. melanogaster. These cell lines were subsequently used to test their response to extracellular adenosine signal by the measurement of cell viability and level of second messengers cAMP and Ca2+ in Cl8+ cells.
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