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Identification of basolateral localization motifs of the urate transporter GLUT9 using functional studies.
Pácalová, Eliška ; Krylov, Vladimír (advisor) ; Pavelcová, Kateřina (referee)
The GLUT9 transporter, coded by the SLC2A9 gene, is one of the key proteins enabling the trasnfer of uric acid across the membrane in epithelial cells of the proximal tubule. In humans, this protein is naturally expressed in two variants: long (GLUT9L) and short (GLUT9S), which differ from one another by their N-terminus sequence. Each of these isoforms is localized on a different pole of the epithelial cell. The signal sequence/motif responsible for this difference is presumed to be located in the aforementioned N-terminus domain. Numerous allelic variants influencing the transport properties of the protein have also been described. The first aim of this thesis is to verify the influence of a newly discovered variant, characterized by substituting of valine for leucine in the 114th position (V114L, in the short form its corresponding variant V85L) on the ability to transport uric acid. Second aim is to verify the influence of mutations in selected motifs, which could be responsible for the localization of the protein, thus also changing its transport properties. Two dileucine motifs 12LGL14 and 33LL34 and one tyrosine motif 84YIKA87 were tested. Functional studies using 14 C radiolabeled urate demonstrated significant decrease of transport ability for the V114L/V85L allelic variant in both...
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Formation of pole assymetry during the Xenopus laevis oocytes growing
Pácalová, Eliška ; Vávra, Jiří (advisor) ; Drutovič, David (referee)
A frog Xenopus laevis is an important model organism used in developmental and cell biology and physiology. Oocytes of this species are frequently used for study of mechanisms of oogenesis and membrane proteins. Through oogenesis an animal-vegetal axis is established by asymmetric distribution of determinant molecules which are important in embryogenesis and establishment for another body axes. The main aim of this thesis is to overview transport mechanisms for establishment of asymmetric molecule (especially mRNA) and organelles asymmetry to animal and vegetal poles and their regulation. Key words: Xenopus laevis, oocyte, animal pole, vegetal pole, cytoskeleton, intracellular transport, mitochondrial cloud, METRO, early pathway, late pathway
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