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Crosstalk of integrin and mTOR signaling
Teglová, Lucie ; Rösel, Daniel (advisor) ; Libus, Jiří (referee)
iv Abstract Crosstalk of integrin and mTOR signalling is an essential process that monitors cellular interaction with extracellular matrix and transmits these inputs to cell growth signalling. Although adhesion status of the cell monitored by integrin signalling is clearly important for regulation of cellular growth, a little is known about the crosstalk of integrin and mTOR signalling. In this study, we employed two different approaches to describe and elucidate character of this crosstalk. p130Cas is an adaptor protein phosphorylated by Src kinase and focal adhesion kinase upon integrin ligand binding and implicated in cell adhesion, motility and survival in both Src-transformed and untransformed cells. Recently, p130Cas was also described in cellular pathology, mainly by its ability to stimulate cell invasion and metastasis. In this study, we described that p130Cas affects mTOR signalling in Src-transformed cells. Substrate domain of p130Cas was found to be indispensable for this effect and is also responsible for serum-induced activation of mTOR signalling. In addition, we prepared cell lines overexpressing various Rheb protein versions and characterized them in context of mTOR signalling, integrin signalling and cell cycle progression. Interestingly, a cell line overexpressing constitutively active...
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Autophagy in yeast
Nejedlý, Adam ; Palková, Zdena (advisor) ; Libus, Jiří (referee)
Autophagy is a process of degradation of the cell cytoplasm, proteins, and organelles. It appears mainly as a response to a lack of nutrients or damage of cell structures and components. Yeast Saccharomyces cerevisiae and also Komagataella phapphi are the main model organisms for the research of autophagy. Autophagy is classified by two main criteria. Firstly, we divide autophagy into macroautophagy and microautophagy. During macroautophagy, the phagophore vesicle, called the autophagosome, is created around sequestrated cargo and then fuses with the vacuole. Microautophagy is a process in which a vacuolar membrane directly surrounds and sequester degraded cargo. During non-selective (bulk) autophagy, a random non- specific portion of cytoplasm is degraded. On the other hand, selective autophagy serves as a pathway for the degradation of specific proteins and organelles. Autophagy research has nowadays a great medical significance thanks to the role of autophagy defects in a wide area of human diseases.
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Heme sensor proteins sensing both heme and CO
Andrlová, Dominika ; Martínková, Markéta (advisor) ; Libus, Jiří (referee)
Heme, a protoporhyrin IX iron complex, is an important component of many proteins necessary for oxygen transfer, storage and activation, as well as for electron transfer. Another group of hemoproteins includes heme sensor proteins. They are either capable of detecting heme itself, which can regulate in turn the sensor function (heme-responsive sensors) or heme forms a binding site for small gas molecules (O2, CO and NO) and the heme-based gas sensors are regulated by these diatomic gases. However, in the case of some proteins their classification is not clear showing a properties of both heme sensor proteins families. Their functions are regulated by heme interaction and a further change in their function after binding of a gas molecule to heme was observed. This summary search is focused on specific representatives of heme-responsive sensors (which function is regulated by heme binding), in which the further influence of the CO molecule on their functions have recently been observed. It is discussed whether some heme-responsive sensors are also heme-based CO sensors aiming the most recent findings about the selected specific heme sensors representatives. Key words: heme, heme sensor proteins, heme-based gas sensors, CO sensors, heme-responsive sensors, heme redox sensors
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Transcriptional response of plants to genotoxic stress and water deficit
Libus, Jiří ; Štorchová, Helena (advisor) ; Cvrčková, Fatima (referee) ; Kovařík, Aleš (referee)
7 Conclusions 1) cDNA array was prepared and used to assay transcript abundances of 376 selected Ara- bidopsis transcripts following various treatments with MMS. a) LoTrEC, clustering algorithm based on local trends of expression profiles, was de- signed and applied to the data. It succeeded to discover functionally rellevant clusters of expression profiles. b) Transcriptional responses to various MMS treatment regimes were investigated. While high MMS concentration seemed only to induce nonspecific stress reaction, the low and combined MMS resulted in a set of more specific expression changes. c) Expression levels of five transcripts were estimated by qRT-PCR. Trends of most of the profiles were confirmed. 2) Expression of 3 genes related to drought stress and/or response to cytokinin were mea- sured by qRT-PCR in wild type and ZOG1 transgenic plants. Transcript levels of all the genes were altered by water deficit. a) Although there are no significant macroscopic differences between wild type and ZOG1 transgenic plants, the mRNA abundances appeared to be influenced by the genotype. b) Leaf position (age) significantly influenced the expression of cig1 and ZOG1 driven by SAG12 promoter. c) RT primed with oligo-dT appeared more eficient than random hexanucleotide-primed reaction for 3 out of 5 mRNAs...
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Crosstalk of integrin and mTOR signaling
Teglová, Lucie ; Libus, Jiří (referee) ; Rösel, Daniel (advisor)
iv Abstract Crosstalk of integrin and mTOR signalling is an essential process that monitors cellular interaction with extracellular matrix and transmits these inputs to cell growth signalling. Although adhesion status of the cell monitored by integrin signalling is clearly important for regulation of cellular growth, a little is known about the crosstalk of integrin and mTOR signalling. In this study, we employed two different approaches to describe and elucidate character of this crosstalk. p130Cas is an adaptor protein phosphorylated by Src kinase and focal adhesion kinase upon integrin ligand binding and implicated in cell adhesion, motility and survival in both Src-transformed and untransformed cells. Recently, p130Cas was also described in cellular pathology, mainly by its ability to stimulate cell invasion and metastasis. In this study, we described that p130Cas affects mTOR signalling in Src-transformed cells. Substrate domain of p130Cas was found to be indispensable for this effect and is also responsible for serum-induced activation of mTOR signalling. In addition, we prepared cell lines overexpressing various Rheb protein versions and characterized them in context of mTOR signalling, integrin signalling and cell cycle progression. Interestingly, a cell line overexpressing constitutively active...
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