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Chromatin immunoprecipitation of selected transcription factors
Smetanová, Jitka ; Vališ, Karel (advisor) ; Převorovský, Martin (referee)
The TEAD family of transcription factors regulates expression of genes affecting cell proliferation, differentiation and apoptosis. The activity of a particular transcription factor called TEAD1 is regulated by the Hippo signalling pathway. The Hippo pathway has been implicated to play a role in cancer suppression, however its precise mechanism remains unclear. MYC and GLUT1, genes which are coding two key regulators of glycolysis, were recently described as potential targets of the Hippo signalling pathway in human leukemia cells. In this diploma thesis, I tried to confirm the proposed interaction of the transcription factor TEAD1 with regulatory sequences of MYC and GLUT1 genes using chromatin immunoprecipitation (ChIP) analysis in human leukemic cells. However, I failed to successfully isolate TEAD1 complexes using ChIP. So, I discuss in my diploma thesis also possible reasons for this outcome, including biological and methodological issues. (In Czech) Key words: Transcriptional regulation, TEAD transcription factors, chromatin immunoprecipitation, leukemia
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Chromatin immunoprecipitation of selected transcription factors
Smetanová, Jitka ; Vališ, Karel (advisor) ; Převorovský, Martin (referee)
The family of transcription factors TEAD regulates the expression of genes that affect cell proliferation, differentiation and apoptosis. Activity of TEAD1 is regulated via the Hippo signaling pathway. General mechanism of tumor cell suppression by the Hippo signaling pathway remains unclear. C-MYC and GLUT1, the two key regulators of glycolysis, were recently described as targets of the Hippo signaling pathway in human leukemia cells. In this diploma thesis, the interaction of TEAD1 with M-CAT binding motifs was experimentally confirmed in the first exon of C-MYC gene. In addition, a new interaction of TEAD1 with M-CAT binding motifs has been found in the enhancer of C-MYC promoter and enhancer of GLUT1 promoter by ChIP analysis. Regulation of glucose metabolism by the Hippo signaling pathway may represent a new mechanism of tumor cell suppression. Key words: Gene regulation, transcription factors, chromatin immunoprecipitation, bioinformatics
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Chromatin immunoprecipitation of selected transcription factors
Smetanová, Jitka ; Vališ, Karel (advisor) ; Převorovský, Martin (referee)
The TEAD family of transcription factors regulates expression of genes affecting cell proliferation, differentiation and apoptosis. The activity of a particular transcription factor called TEAD1 is regulated by the Hippo signalling pathway. The Hippo pathway has been implicated to play a role in cancer suppression, however its precise mechanism remains unclear. MYC and GLUT1, genes which are coding two key regulators of glycolysis, were recently described as potential targets of the Hippo signalling pathway in human leukemia cells. In this diploma thesis, I tried to confirm the proposed interaction of the transcription factor TEAD1 with regulatory sequences of MYC and GLUT1 genes using chromatin immunoprecipitation (ChIP) analysis in human leukemic cells. However, I failed to successfully isolate TEAD1 complexes using ChIP. So, I discuss in my diploma thesis also possible reasons for this outcome, including biological and methodological issues. (In Czech) Key words: Transcriptional regulation, TEAD transcription factors, chromatin immunoprecipitation, leukemia
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Chromatin immunoprecipitation of selected transcription factors
Smetanová, Jitka ; Vališ, Karel (advisor) ; Převorovský, Martin (referee)
The family of transcription factors TEAD regulates the expression of genes that affect cell proliferation, differentiation and apoptosis. Activity of TEAD1 is regulated via the Hippo signaling pathway. General mechanism of tumor cell suppression by the Hippo signaling pathway remains unclear. C-MYC and GLUT1, the two key regulators of glycolysis, were recently described as targets of the Hippo signaling pathway in human leukemia cells. In this diploma thesis, the interaction of TEAD1 with M-CAT binding motifs was experimentally confirmed in the first exon of C-MYC gene. In addition, a new interaction of TEAD1 with M-CAT binding motifs has been found in the enhancer of C-MYC promoter and enhancer of GLUT1 promoter by ChIP analysis. Regulation of glucose metabolism by the Hippo signaling pathway may represent a new mechanism of tumor cell suppression. Key words: Gene regulation, transcription factors, chromatin immunoprecipitation, bioinformatics
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Molecular mechanism of MST1 kinase activation in cancer cells
Smetanová, Jitka ; Vališ, Karel (advisor) ; Grobárová, Valéria (referee)
MST1 kinase is an internal part of the Hippo signal pathway. The Hippo pathway is an evolutionary conserved regulator of tissue and organ growth and affects proliferation and apoptosis. Active MST1 kinase phosphorylates YAP and TAZ oncoproteins, which regulate the activity of transcription factors in their unphosphorylated state, including TEAD and SMAD. Furthermore active MST1 kinase phosphorylates FOXO transcription factors and induces their translocation into the cell nucleus. Finally the activation of MST1 kinase leads to cell apoptosis or halt cell cycle in G1 phase. Activation of MST1 protein depends on its auto-phosphorylation and cleavage. Recently, there are several articles which take interest in the issue of activation of MST1. Some of them describe the activation of MST1 by the effector caspase-3 and -7, on the other hand the latest articles argue that MST1 kinase itself is responsible for the activation of caspases. The molecular mechanism of MST1 kinase activation was studied in this bachelor thesis. We used the biologically active compounds GDC-0941 for the activation of MST1 protein. The activity of caspase was inhibited by specific inhibitor Z-DEVD. Using electrophoresis and Western blot it was demonstrated that MST1 is active in the case when caspases are inhibited. This fact...
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Economic Relations between the Czech Republic and the Republic of South Africa
Smetanová, Jitka ; Zamykalová, Miroslava (advisor) ; Chládek, Miloslav (referee)
This work analyses the relations of the Czech Republic and South Africa in the field of trade and investment. The first chapter describes the environment of South Africa from geographic, demographic, historical, political views and perspectives of involvement in various organizations. The second chapter compares the economic environments in the Czech Republic and South Africa in the fields of macroeconomic indicators, foreign trade and foreign direct investment. The third chapter deals with mutual economic relations between the two countries. The fourth chapter focuses on the company Crystalex CZ dealing with South African partners.
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