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Human 4E protein family in stress granules granules and their further characterization
Hrbková, Pavlína ; Frydrýšková, Klára (advisor) ; Hašek, Jiří (referee)
Eukaryotic initiation factor 4E (eIF4E) is a key part of initiation and regulation of translation in human cells. Three members of human eIF4E proteins have been characterized: eIF4E1, eIF4E2 and eIF4E3. Cellular stress causes translation initiation inhibition followed by disassembly of the polysomes, those processes are accompanied by the assembly of cytoplasmic RNA granules, called stress granules (SG). Stress granules are dynamic structures whose composition may vary depending on the cell type and the stress stimulus. In this study, human cells were subjected to the following stress conditions: high temperature (HS), sodium arsenite (AS) or hypoxia. Using fluorescence microscopy, pairs of human translational initiation factors from the 4E protein family were visualized and their localization to SG was assessed with one GFP- 4E incorporated in the stable cell line and the other one detected endogenously. Here we show eIF4E1 being a part of all the SGs, both in HS and AS conditions. Next, the eIF4E1 and eIF4E3 proteins together form more SGs than proteins eIF4E1, respectively eIF4E3, with eIF4E2. And last, that the presence of the particular 4E protein has no effect on the composition of SGs. Furthermore, selected groups of proteins were assessed for their potential to localize to the SGs under HS...
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Yeast colonies as a model of multicellular behaviour of microorganisms
Kuthan, Martin ; Palková, Zdena (advisor) ; Hašek, Jiří (referee) ; Kolarov, Jordan (referee)
- 32 - Závěr Již v úvodu jsem se zmínil, že mnohé laboratorní kmeny kvasinek ztratili vlivem domestikace schopnost filamentálního růstu. Ukázkovým příkladem cílené domestikace je laboratorní kmen S288C. Historie vzniku tohoto kmene je zajímavá a dokumentuje výrazný vliv genetiků na jeho vlastnosti. Velké množství kmenů S. cerevisiae používaných v laboratořích má společného předka kterým je kmen EM93. Tento diploidní kmen izoloval v Kalifornii v roce 1938 Emil Mrak z hnijícího fíku (MORTIMER and JOHNSTON 1986). Není však jasné, zda se jednalo o přirozenou mikroflóru fíků, nebo o kontaminaci komerčními pekařskými či kvasnými kmeny. Kmen EM93 je heterothalický a je schopen filamentálního růstu (LIU et al. 1996). Pro laboratorní účely ale nebyl úplně ʺpohodlnýʺ. Protože tvorba shluků buněk brání izolaci klonů vzniklých z jediné buňky a ztěžuje přesné určení počtu buněk v tekutých kulturách, snažili se genetici získat prototrofní kmen s neadherujícími, dobře resuspendovatelnými buňkami. Mnohonásobným křížením haploidních segregantů kmene EM93 s dalšími laboratorními kmeny a komerčními pekařskými kmeny vytvořil Robert Mortimer kmen S288C, jehož 90 % genomu pochází...
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The role of plasma membrane Na+/H+ antiporters in the physiology of yeast cells
Papoušková, Klára ; Sychrová, Hana (advisor) ; Hašek, Jiří (referee) ; Flachs, Pavel (referee)
) r Nhal-type antiporterswith differentsubstratespeciÍicity(or affinityto individualsubstrates) andfunctions.One of themis involvedin thedetoxificationof cells andtheothercouldplay an importantrole in the maintenanceof stableintracellularpotassiumcontent,cell volume andpH of thecýoplasmthanlsto its abilitytotransportK+. At this time, no yeast speciespossessingsolely a Na*/fI*antiporterwith narrow substratespecificity (for Na* and Li) is known;therefore,the physiologicalrole of Nhal- type antiportersseemsto be complexwith a participationboth in the eliminationof toxic cationsfromcells andin themaintenanceof stableintracellularK* concentration,cell volume andcýoplasmic pH. 4.Summary Themainresultsof thethesíscanbesummarizedas: o The studiesof sequencesof yeastalkali metalcation/Iťantiporters(Nhal, Nhxl andKhal families)in silico showedthatthe structureof all threegroupsof theseproteinsprobably contains 12 transmembranesegments,we found some conservedamino acid residuesor motifslikely to be importantforproperfunctioningof antiporters,andourphylogeneticstudy revealedthatNhxl proteinsare relatedto both plant and mammalianNa*/Iť antiporters, Khal antiportersare close to thebacterialrelativesandNhal-type proteinsform a soparate goup so far characteizedonly in yeastsor fungi. o The D. hanseniiNHAI geneencodesa...
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Regulatory mechanisms of microtubule reorganization in activated mast cells
Rubíková, Zuzana ; Dráber, Pavel (advisor) ; Binarová, Pavla (referee) ; Hašek, Jiří (referee)
Microtubules (MTs) are highly dynamic structures essential for the spatio-temporal intracellular organization and transport, signal propagation, cell differentiation, motility and division. To perform these roles, MTs create arrangements capable of fast and precise adaptation to various signals. MTs are under the control of many factors regulating MT nucleation, stability and dynamics. Bone marrow-derived mast cells (BMMCs) are important immune system cells, which can cause serious diseases if their functions are deregulated. Although MT reorganization during BMMC activation is well established, the molecular mechanisms that control their remodelling are largely unknown. In the presented thesis we functionally characterised GIT1/βPIX signalling proteins, PAK1 kinase, and Ca2+ signalling in the regulation of MT nucleation in BMMCs and other cell types. We also elucidated the function of miltefosine (hexadecylphosphocholine), a promising candidate for the treatment of mast cell-driven diseases. We found that GIT1/βPIX signalling proteins are γ-tubulin-interacting proteins associating with centrosomes in BMMCs. MT nucleation is positively regulated by GIT1 and Ca2+ , whereas βPIX is a negative regulator of MT nucleation in BMMCs. Cytosolic Ca2+ affects γ-tubulin properties and stimulates the...
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Management of the Construction Project
Hašek, Jiří ; Rusek, Jan (referee) ; Nováková, Jana (advisor)
The bachelor thesis deals with the issue of construction project management and explains the basic concepts that concern this area. The aim of this work is to get acquiainted with the basic methods of project management and their aplication on concrete practical project.
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