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The role of SNW/SKIP proteins in splicing
Hollá, Sandra ; Novotný, Marian (advisor) ; Kozáková, Eva (referee)
SNW proteins are essential nuclear factors whose name was derived from the conserved motif SNW. They can be found with various representatives across the eukaryotic empire. SNW proteins are involved in regulating many cellular processes - for example regulation of gene expression and cell cycle. By far the most information about the function of human homologue provides SKIP/NCoA-62, which was found in a number of signaling pathways. Yet it is not fully understood the function of SNW proteins. In this work, I would like to focus on the role of SNW proteins in pre-mRNA splicing, particularly in the yeast Saccharomyces cerevisiae, where most results were acquired. Key words: SNW protein, Prp45, splicing pre-mRNA, SKIP/NCoA-62, Prp22
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Mechanisms of invasiveness and transcription regulation in cancer cells
Tolde, Ondřej ; Folk, Petr (advisor) ; Kovář, Jan (referee) ; Brdička, Tomáš (referee)
The mechanisms of invazivity and regulation of transcription of cancer cells Cancer originates in cells that overcome the control mechanisms of the organism. Cancer cells can be eventually released from the site of origin and spread through tissues. Cancer cells can acquire certain mechanisms that enable them to more effectively invade surrounding tissue or layers of other cells. The research on the migration of cancer cells is important for the understanding of the origin and spreading of metastases and consequently for anticancer therapy. In my Ph.D. work, I participated in the research of the properties of invasive metastatic cells. We compared non-invasive rat sarcoma cell line with a higly metastatic cell line derived from it. We showed that cells of the invasive cell line use amoeboid mode of migration, have upregulated Rho/ROCK signaling, and have accumulated actin and myosin at the leading edge. It is at the leading edge where the cells generate their traction forces. Cells of non-invasive cell line use mesenchymal mode of migration and generate forces mainly at their retracting end. We also compared two breast cancer cell lines derived from a single carcinoma. We showed that the more invasive cell line, derived from its parental line by neoplastic transformation, displayed elevated cytoskeletal...
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The role of evolutionarily conserved proteins BIR-1/Survivin and SKP-1 in the regulation of gene expression
Kostrouch, David ; Kostrouch, Zdeněk (advisor) ; Dráber, Pavel (referee) ; Pacák, Karel (referee)
SKIP and BIR/Survivin are evolutionarily conserved proteins. SKIP is a known transcription and splicing cofactor while BIR-1/Survivin regulates cell division, gene expression and development. Loss of function of C. elegans SKIP (SKP-1) and BIR-1 induces overlapping developmental phenotypes. In order to uncover the possible interactions of SKP-1 and BIR-1 on the protein level, we screened the complete C. elegans mRNA library using the yeast two-hybrid system. These experiments identified partially overlapping categories of proteins as SKP-1 and BIR-1 interactors. The interacting proteins included ribosomal proteins, transcription factors, translation factors and cytoskeletal and motor proteins suggesting involvement of the two studied proteins in multiple protein complexes. To visualize the effect of BIR-1 on the proteome of C. elegans we induced a short time pulse BIR-1 overexpression in synchronized L1 larvae. This led to a dramatic alteration of the whole proteome pattern indicating that BIR-1 alone has the capacity to alter the chromatographic profile of many target proteins including proteins found to be interactors in yeast two hybrid screens. The results were validated for ribosomal proteins RPS-3, RPL-5, non-muscle myosin and TAC-1, a transcription cofactor and a centrosome associated...
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Library regional educatuional centers (activity analysis)
Kniezková, Michaela ; Černá, Milena (advisor) ; Stöcklová, Anna (referee)
The thesis work deals with the system of linking-up education of librarians, which is implemented through the region's education centres which work in the county libraries.In this centre there is National Library of The Czech Republic, which in the partnership of the professional institutions digestedly operates and coordinates the function of the provided services which are operate by regional centres.The target is to provide relevant information, which might be use while framing the education activities of librarian regional centres.The part of my work is the research aimed to analyze of the partial implications, which might effect the quality and the enforcement of its function.
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Splicing of atypical introns in S. cerevisiae
Cit, Zdeněk ; Půta, František (advisor) ; Pichová, Alena (referee)
Pre-mRNA splicing is a vital process of gene expression important for all eukaryotic organisms. For the proper function of this very complex and dynamic event the presence of few specialized RNA and many proteins that hold a variety of tasks is necessary, not only inside the splicing complex itself, but also beyond this complex. The Prp45 is one of the proteins involved in pre-mRNA splicing in yeast Saccharomyces cerevisiae. Its human homologue, SNW1/SKIP, is involved in splicing but also in other crucial cell processes. The Prp45 protein was reliably reported only to participate in the second transesterification reaction of splicing. But there are also data suggesting its possible involvement in the first transesterification reaction. This work provides further evidences linking protein Prp45 with the first splicing reaction, obtained by the research of cells carrying the mutant allele prp45(1-169). Cells carrying this allele show dropped splicing and accumulation of pre-mRNAs. This thesis therefore also investigated the possible influence of Prp45 protein on the RNA export from the nucleus to the cytoplasm. But no connection between this protein and RNA transport was discovered. Keywords pre-mRNA splicing; Saccharomyces cerevisiae; Prp45; Mer1; Mud2; Prp22; Rrp6; AMA1; SNW1/SKIP
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Notch-independent functions of CSL transcription factors
Teska, Mikoláš ; Folk, Petr (advisor) ; Převorovský, Martin (referee) ; Kuthan, Martin (referee)
Notch pathway plays a critical role during development and life of Metazoan organisms. CBF1 is a component of the Notch pathway that mediates the regulation of target genes. The discovery of CBF1-like proteins in yeast raised the question of their function in unicellular organisms - before the origin of canonical Notch pathway. CBF1-homologs in yeast are conserved in parts that are important for DNA binding and bind to CBF1-binding elements in vitro. CBF1 and related transcription factors in Metazoa (CSL) interact with many proteins in Notch-dependent as well as Notch-independent complexes. The Notch receptor has likewise some CSL-independent functions. This assay reports about interacting partners of CSL in Metazoa along with homologous proteins in yeast with the aim to highlight potential interactions of CBF1-homologs in evolutionary ancestral context.
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The role of SNW/SKIP proteins in splicing
Hollá, Sandra ; Novotný, Marian (advisor) ; Kozáková, Eva (referee)
SNW proteins are essential nuclear factors whose name was derived from the conserved motif SNW. They can be found with various representatives across the eukaryotic empire. SNW proteins are involved in regulating many cellular processes - for example regulation of gene expression and cell cycle. By far the most information about the function of human homologue provides SKIP/NCoA-62, which was found in a number of signaling pathways. Yet it is not fully understood the function of SNW proteins. In this work, I would like to focus on the role of SNW proteins in pre-mRNA splicing, particularly in the yeast Saccharomyces cerevisiae, where most results were acquired. Key words: SNW protein, Prp45, splicing pre-mRNA, SKIP/NCoA-62, Prp22
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Mechanisms of invasiveness and transcription regulation in cancer cells
Tolde, Ondřej ; Folk, Petr (advisor) ; Kovář, Jan (referee) ; Brdička, Tomáš (referee)
The mechanisms of invazivity and regulation of transcription of cancer cells Cancer originates in cells that overcome the control mechanisms of the organism. Cancer cells can be eventually released from the site of origin and spread through tissues. Cancer cells can acquire certain mechanisms that enable them to more effectively invade surrounding tissue or layers of other cells. The research on the migration of cancer cells is important for the understanding of the origin and spreading of metastases and consequently for anticancer therapy. In my Ph.D. work, I participated in the research of the properties of invasive metastatic cells. We compared non-invasive rat sarcoma cell line with a higly metastatic cell line derived from it. We showed that cells of the invasive cell line use amoeboid mode of migration, have upregulated Rho/ROCK signaling, and have accumulated actin and myosin at the leading edge. It is at the leading edge where the cells generate their traction forces. Cells of non-invasive cell line use mesenchymal mode of migration and generate forces mainly at their retracting end. We also compared two breast cancer cell lines derived from a single carcinoma. We showed that the more invasive cell line, derived from its parental line by neoplastic transformation, displayed elevated cytoskeletal...
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The role of evolutionarily conserved proteins BIR-1/Survivin and SKP-1 in the regulation of gene expression
Kostrouch, David ; Kostrouch, Zdeněk (advisor) ; Dráber, Pavel (referee) ; Pacák, Karel (referee)
SKIP and BIR/Survivin are evolutionarily conserved proteins. SKIP is a known transcription and splicing cofactor while BIR-1/Survivin regulates cell division, gene expression and development. Loss of function of C. elegans SKIP (SKP-1) and BIR-1 induces overlapping developmental phenotypes. In order to uncover the possible interactions of SKP-1 and BIR-1 on the protein level, we screened the complete C. elegans mRNA library using the yeast two-hybrid system. These experiments identified partially overlapping categories of proteins as SKP-1 and BIR-1 interactors. The interacting proteins included ribosomal proteins, transcription factors, translation factors and cytoskeletal and motor proteins suggesting involvement of the two studied proteins in multiple protein complexes. To visualize the effect of BIR-1 on the proteome of C. elegans we induced a short time pulse BIR-1 overexpression in synchronized L1 larvae. This led to a dramatic alteration of the whole proteome pattern indicating that BIR-1 alone has the capacity to alter the chromatographic profile of many target proteins including proteins found to be interactors in yeast two hybrid screens. The results were validated for ribosomal proteins RPS-3, RPL-5, non-muscle myosin and TAC-1, a transcription cofactor and a centrosome associated...
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