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Functional characterization of LACE1 APTase and mitochondrial AAA proteases YME1L and AFG3L2 in mitochondrial protein homeostasis.
Tesařová, Jana ; Stibůrek, Lukáš (advisor) ; Kalous, Martin (referee) ; Pecina, Petr (referee)
Mitochondrial protein homeostasis is crucial for cellular function and integrity. It is ensured by many specific mitochondrial proteases with possible chaperone functions located across the various mitochondrial subcompartments. In the first part, we have focused on characterization of functional overlap and cooperativity of proteolytic subunits AFG3L2 and YME1L of the mitochondrial inner membrane complexes m- and i-AAA in HEK293 cells. The double AFG3L2/YME1L knockdown cells showed severe alteration in OPA1 protein processing, marked elevation in OMA1 protease and severe reduction in SPG7. Our results reveal cooperative and partly redundant involvement of AFG3L2 and YME1L in the maintenance of mitochondrial protein homeostasis and further emphasize their importance for mitochondrial and cellular function and integrity. The aim of the second part was to characterize the cellular function of LACE1 (lactation elevated 1) in mitochondrial protein homeostasis. LACE1 protein is a human homologue of yeast Afg1 (ATPase family gene 1) ATPase. We show that LACE1 is a mitochondrial integral membrane protein that exists as a part of three complexes of approximately 140, 400 and 500 kDa. We demonstrate that LACE1 mediates degradation of nuclear-encoded complex IV subunits COX4, COX5A and COX6A. Using affinity...
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Mechanisms and aplications of macromolecule translocation across membranes of eukaryotic cells by bacterial toxins
Poledňák, Jan ; Fišer, Radovan (advisor) ; Žáčková Suchanová, Jiřina (referee)
Toxin translocation across the cytoplasmic membrane of the eukaryotic cell is a potent virulence factor of bacteria causing disease to eukaryotic organisms. Toxins translocate their domains responsible for the toxic activity inside the cell or create pores in cell membrane allowing the transmembrane traffic of ions, DNA, RNA or proteins. Knowledge of the toxin translocation process enables to characterize the mechanism and also the properties of the pore-forming toxin. Some of these toxins have been described in such a detail that were changed using site-directed mutagenesis and can serve as tools for characterization of the translocated molecules. One of such examples is the transfer of nucleotides or the whole nucleic acid molecules across the membrane through the pore of S. aureus α-hemolysine. Nowadays, this application is commercially used for DNA sequencing. Keywords: translocation, bacterial toxins, plasmatic membrane, nanopore sequencing
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Mechanisms and aplications of macromolecule translocation across membranes of eukaryotic cells by bacterial toxins
Poledňák, Jan ; Fišer, Radovan (advisor) ; Žáčková Suchanová, Jiřina (referee)
The bacterial protein toxins endowed with the ability to translocate across the plasmatic membrane are often crucial virulence factors of pathogenic bacteria invading eukaryotic organisms. These toxins translocate either their own protein domains carrying toxic activity or can form pores transferring other substances like small ions, DNA, RNA or proteins. By observing the translocation of these molecules together with other artificially prepared agents on synthetic membranes it allows detailed understanding of mode of action of individual pore-forming toxins. Some of the toxins were actually described in such a detail that can serve as tools for drug delivery or characterization of new translocated molecules. One of such examples is the transfer of nucleotides or the whole nucleic acid molecules across the membrane pore of S. aureus α-hemolysine. Nowadays, this application is commercially used for DNA sequencing. Keywords: translocation, bacterial toxins, plasmatic membrane, nanopore sequencing
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The role of trehalose in mycorrhizal associations
Šoch, Jan ; Ponert, Jan (advisor) ; Hála, Michal (referee)
Mycorrhizal symbiosis is a widely spread phenomenon in nature. A translocation of nutrients occurs between symbionts with disaccharide trehalose playing a key role in the process. However, this saccharide fulfils many important roles in metabolism of fungi and plants. Fungi use trehalose mainly as storage and transport saccharide. On the other hand, trehalose occurs in extremely low amounts in plants where it acts as a signal molecule. Thus it is likely that the saccharide could be used by the fungus to manipulate plant metabolism on a physical interface between symbionts. Trehalose has a similar function in many parasitic interactions. In most cases of mycorrhizal associations trehalose synthesis creates a carbon sink in mycelium which leads to saccharide transfer from the host plant to the fungus. Completely different situation occurs in the types of mycorrhizal symbiosis, where saccharides are translocated from a fungus to a plant. Some plants can utilize trehalose effectively as a sole source of energy. Consequently, the question raises - could such plants gain fungal trehalose on purpose as a source of energy and carbon? This review aims to assess and discuss the mentioned possibilities considering available literature. Key words: mycorrhiza, orchids, parazitism, saccharides, sink, symbiosis,...
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Preparation and phytoextraction of 125-I labelled pharmaceuticals
Luptáková, Dominika ; Smrček, Stanislav (advisor) ; Soudek, Petr (referee)
Pharmaceuticals are group of organic substances with significant worldwide consumption in human and veterinary medicine. These compounds may be metabolized in the organism, but in some cases they remain unchanged and both are usually excreted via renal excretion in the native form or as metabolites. Large quantities of pharmaceuticals and their metabolites contaminate municipal wastewater. The wastewater treatment plants are unable to remove these substances completely, so they contaminate surface water, groundwater and soil as well. Due to the biological activity of pharmaceuticals, long - term effect may cause bacterial resistance, endocrine influence, DNA and renal damages in non-target organisms. The phytoextraction and the translocation of radiolabeled diclofenac with 125 I were experimentally studied by using of in vitro cultivated plants Helianthus annuus and Zea mays. Efficiency od phytoextraction was monitored as decrease of radioactivity of tested substance [125 I]diclofenac in Murashige-Skoog cultivation medium. Both species are able to extract tested substance during 8 to 10 days of cultivation, with efficiency approximately 85 % using Zea mays and 79 % using Helianthus annuus. Better extraction ability of diclofenac was observed at Helianthus annuus - 80 mg/ kg of dry weight compared...
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Integration of clonal plants in heterogeneous environment
Vinšová, Jana ; Weiser, Martin (advisor) ; Macek, Petr (referee)
Clonal plants may be able to cope with spatial and temporal heterogeneity due to the physiological integration of ramets. To begin with, I discuss basic characteristics of clonal plants and then I provide a review of theoretical and empirical work dealing with the advantages and disadvantages of clonal growth in heterogenous conditions. Three main traits - translocation of resources, division of labour and foraging - are examined in terms of their contribution to the success of clonal plants. Despite a large amount of attention which this topic has received in the past few decades, a number of issues still remain unclear.
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The process of translocation in Ostrov nad Ohří (Schlackenwerth). The origin and development of the church of St Jacob and the church of St Nicolas according to the urbanization
Šídlová, Andrea ; Scholz, Stefan (advisor) ; Záruba, František (referee)
The thesis should notice the time when the two churches arose by looking back at the historical situation in the 13th century Premyslid Bohemia. Furthermore, we will try to evaluate the social, legal and property situation of the given area, and it will monitor the development and building. The work is given a chapter on the administrative area of Sedlecko to point out conflicts of interest of Czech kings and Roman rulers in this territory and the formation of the state historic town of Cheb and between the 12th and 13th century. The origins of Ostrov agglomeration is linked to the family of Hrabišic. The following chapter discusses their family, their asset base and foundation. The main part is devoted to the translocation and the relocation of settlements from the center of the parish church of St. James to St. Nicholas. The reason was, as it was previously shown, an unsatisfactory position of the original settlement for its further development, which was necessary for the competitiveness of the local market center. It subsequently evolved into a town. This situation culminated in the privilege of King John of Luxembourg. At that time a new and relatively big church was being built which replaced the church of St. James. It was a part of the settlement and farmstead of the family of Hrabišic and...
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Significance of MLL gene aberrations in patients with acute myeloid leukemia
Šárová, Iveta
In acute myeloid leukemia (AML), predominantly in AML M5a, the most frequent recurrent aberration of chromosome 11 involves region 11q23. Molecular breakpoint studies of several translocations involving chromosomal band 11q23 led to the detection of a gene that was named MLL (myeloid/lymphoid leukemia). This gene is important for the proper HOX gene expression during ontogenesis and hematopoiesis. Chromosomal aberrations affecting the MLL gene occur in 5 - 10 % of AML cases and are very variable. Since that time, more than 70 different translocation partners of the MLL gene have been described. Aberrations of the MLL gene are associated with an aggresive type of the disease and its detection is needed for the treatment decision. Therefore, we investigated the occurrence of MLL abnormalities in bone marrow cells of the 66 newly diagnosed AML patients, using conventional cytogenetic and fluorescence in situ hybridization (FISH) analyses with a commercially available MLL Break Apart Rearrangement probe (Abbott VYSIS). Out of the 66 patients, we proved MLL abnormalities in 9 (13,6%): 5 (7,6%) showed translocation of MLL gene, in 3 (4,5%) we detected MLL gene amplification without any evidence of rearrangement and in 1 (1,5%) pacient only an extra copy of the MLL gene. The FISH results were verified by...
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