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Crosstalks between nitrogen metabolism, the TOR pathway and the metabolism of lipids
Grulyová, Michaela ; Převorovský, Martin (advisor) ; Maršíková, Jana (referee)
Cells coordinate their metabolism based on various factors, for example nitrogen availability. The TOR pathway is an important regulator of nitrogen metabolism, it has a role in sensing intracellular amino acids status, and it controls especially cell growth, protein synthesis, proliferation and cell survival. However, it has been shown that the TOR pathway also controls lipid biosynthesis and lipid accumulation through various mechanisms in response to nitrogen availability. Although the TOR pathway is well conserved among the eukaryotic organisms, its outcomes differ diametrically when it comes to the lipid accumulation. This essay provides some insides into the mechanisms of regulation of the lipid metabolism by the TOR pathway.
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Glutamine synthetases and their role in nitrogen metabolism in plant shoot
Kobercová, Eliška ; Fischer, Lukáš (advisor) ; Tylová, Edita (referee)
Glutamine synthetase is a key enzyme for ammonium assimilation and glutamine biosynthesis in plants. Ammonium ions are important intermediates in nitrogen metabolism, but their elevated concentration is toxic to plants. Correct function of glutamine synthetase is essential to plant life - glufosinate, a specific inhibitor of glutamine synthetase acts as a total herbicide. Glutamine synthetase is involved in a number of important metabolic processes: primary assimilation of nitrogen nutrients, in reassimilation of ammonium ions released during photorespiration or metabolism of phenylpropanoids and in nitrogen remobilization in developing seeds, during germination or senescence. According to localization in the cell glutamine synthetases in angiosperms are divided into the plastid form (GS2), which is typically encoded by a single gene, and the cytosolic form (GS1), which is encoded by a small multigene family. The various isoforms of glutamine synthetases have different location within plant organs and tissues, ways of regulation and role in nitrogen metabolism. Keywords: plastid and cytosolic glutamine synthetase, ammonium assimilation, nitrogen metabolism, photorespiration
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Candida parapsilosis secreted aspartic proteinases: processing and secretion
Vinterová, Zuzana ; Heidingsfeld, Olga (advisor) ; Hodek, Petr (referee) ; Szotáková, Barbora (referee)
Candida parapsilosis is an emerging human opportunistic pathogen causing a wide spectrum of potentially life-threatening infections in immunocompromised hosts. One of the most important virulence factors of Candida spp. is a production of secreted aspartic proteinases (Saps). Presented thesis is mainly focused on the study of secreted aspartic proteinase 1 (Sapp1p) of C. parapsilosis, its processing and secretion under variable conditions and by use of various experimental models. Sapp1p is secreted by C. parapsilosis cells into the extracellular space as a completely processed and fully active enzyme. Experiments studying the C. parapsilosis cell wall (CW) confirmed the prolonged presence of completely processed Sapp1p on the cell surface (CW- Sapp1p). Proteolytic activity assay performed with the intact cells showed that CW-Sapp1p is proteolytically active prior to its release into the extracellular space and is capable of substrate cleavage. Biotinylation experiments with consecutive MS analysis revealed that CW-Sapp1p biotinylation is incomplete but saturable process, leaving partially unlabelled molecules. The accessibility of individual lysine residues in the Sapp1p molecule varied, with exception of four residues that were labelled in all of our experiments performed. The final step of...
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The effect of biotic stress on nitrogen metabolism in tobacco plants
Fiala, Martin ; Ryšlavá, Helena (advisor) ; Müller, Karel (referee)
In this project the effect of viral infection on the metabolism of nitrogenous compounds in tobacco plants (Nicotiana tabacum L. cv. Petit Havana SR1) was studied. The tobacco plants were infected with Potato virus Y, strain NTN, ELISA confirmed the presence of the virus. Enzymes that participate in C4 plants in Hatch-Slack cycle fosfoenolpyruvate carboxylase (EC 4.1.1.31, PEPC), NADP-dependent malic enzyme (EC 1.1.1.40, NADP-ME), pyruvate, phosphate dikinase (EC 2.7.9.1, PPDK) are present also in C3 plants and are related to plant responses to stress conditions. As a result of viral infection, the activities of all these enzymes were increased. Infection caused by PVYNTN decreased activity of nitrate reductase (EC 1.7.1.1, NR), an enzyme catalyzing reduction of nitrates to nitrites. The activity of enzymes catalyzing the synthesis of glutamine from glutamate and ammonium ions: glutamine synthetase (EC 6.3.2.1, GS) and glutamate synthase (EC 1.4.1.14, GOGAT) was enhanced. In addition to this main route of nitrogen fixation the plant can still use glutamate dehydrogenase (EC 1.4.1.2, GDH). This enzyme can also catalyze the opposite reaction, deamination of glutamate. The direction of response depends on environmental conditions. In this case a significant increase of oxidative-deaminating activity...
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