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Regulation of mycobacterial transcription
Kafka, Vojtěch ; Krásný, Libor (advisor) ; Dostálová, Hana (referee)
RNA polymerase (RNAP) is the enzyme that catalyzes synthesis of RNA. Mycobacterial RNAP significantly differs from RNAPs from other bacterial species. It requires special transcription factors such as RbpA or CarD. Another difference is the presence of a small RNA (sRNA), Ms1, that binds to mycobacterial RNAP. Ms1 regulates the amount of RNAP in the cell. In our laboratory we recently discovered MoaB2, a new binding partner of mycobacterial A (encoded by sigA), an RNAP subunit, which is essential for recognition of the initial promoter sequence and initiation of transcription. The function of MoaB2 in the regulation of transcription and gene expression is still unknown. The first aim of this Thesis is contribute to elucidation of the mechanism by which Ms1 affects the amount of RNAP. The experiments revealed that this regulation occurs at the level of transcription; Ms1 affects the activity of promoter(s) that drive the transcription of rpoB- rpoC that encode the two catalytic subunits of RNAP. The second aim of this Thesis is to characterize the interactions of MoaB2 with protein of the transcription apparatus. The results confirmed the interaction of MoaB2 with A and showed that neither RNAP nor transcription factors RbpA and CarD are required for this interaction. Finally, a role of the...
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Regulation of mycobacterial transcription
Kafka, Vojtěch ; Krásný, Libor (advisor) ; Dostálová, Hana (referee)
RNA polymerase (RNAP) is the enzyme that catalyzes synthesis of RNA. Mycobacterial RNAP significantly differs from RNAPs from other bacterial species. It requires special transcription factors such as RbpA or CarD. Another difference is the presence of a small RNA (sRNA), Ms1, that binds to mycobacterial RNAP. Ms1 regulates the amount of RNAP in the cell. In our laboratory we recently discovered MoaB2, a new binding partner of mycobacterial A (encoded by sigA), an RNAP subunit, which is essential for recognition of the initial promoter sequence and initiation of transcription. The function of MoaB2 in the regulation of transcription and gene expression is still unknown. The first aim of this Thesis is contribute to elucidation of the mechanism by which Ms1 affects the amount of RNAP. The experiments revealed that this regulation occurs at the level of transcription; Ms1 affects the activity of promoter(s) that drive the transcription of rpoB- rpoC that encode the two catalytic subunits of RNAP. The second aim of this Thesis is to characterize the interactions of MoaB2 with protein of the transcription apparatus. The results confirmed the interaction of MoaB2 with A and showed that neither RNAP nor transcription factors RbpA and CarD are required for this interaction. Finally, a role of the...
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Function of stress sigma factors of RNA polymerase SigD, SigE, SigH and SigM in transcription regulation network of Corynebacterium glutamicum
Dostálová, Hana ; Pátek, Miroslav (advisor) ; Bobek, Jan (referee) ; Halgašová, Nora (referee)
Grampositive bacterium Corynebacterium glutamicum is an important industrial producer of amido acids and other metabolites. Its genome encodes 7 sigma (σ) subunits of RNA polymerase: primary factor σA , primary-like σB and five alternative sigma factors, σC , σD , σE , σH and σM (sigma factors with extracytoplasmic function). This study is focused on revealing so far unknown regulons of stress sigma factors or closer description of regulons whose genes are controled by σD , σE , σH and σM . These factors were partially described for their activity during surface (σD and σE ), heat (σE , σH and σM ) and oxidative (σH and σM ) stress response. We assumed that the genes of each regulon are transcribed from promoters of a single class. For the purpose of detailed promoter analysis, it was necessary to develop methods which can quickly and reliably assign sigma factor to particular promoters and, thus, respective genes. For this purpose, a combination of in vivo (two-plasmid system) and in vitro (in vitro transcription) techniques was developed that allow to specify this assignment. We identified 9 σH /σE - promiscuous promoters (PamtR, Pcg0378, Pcg1121, Pcg3309, Pcg3344, PclgR, PdnaJ, PdnaK and PsigB), 7 σD /σH - promiscuous promoters (Pcg0607, Pcg2047, Pcmt2, PfadD2, Plpd, PlppS and PrsdA) a 9 σH /σM...
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Methods of investigation of murders : (selected issues)
Dostálová, Hana ; Konrád, Zdeněk (advisor) ; Musil, Jan (referee)
The theme of the thesis is the Methodology of Investigation of Murders. According to the scope of that theme the author concentrated only on selected issues. The first chapter includes the overview of the Czech and foreign literature related to the theme of the thesis. The second chapter covers some of the legal aspects of crimes against live, as they are enumerated in the Czech Criminal Code. The purpose of this chapter is to emphasize the link between the investigation of muders as a criminalistic discipline and the criminal law. This chapter concerns crimes as murder, manslaughter, infanticide, killing of negligence and participation on suicide, including current statistical data with regard to the quantity of murders within the area of the Czech Republic. The third chapter concerns the criminalistic characteristic of the murder, which consists of the modus operandi, the character of the perpetrator, the character of the victim and the motive. The next chapters follow the specifics of particular phases of the metodology of investigation of murders, as they are described in the leading Czech professional literature, concretely typical traces, typical investigation situations, specifics of the object of the investigation and specifics of the impetuses for investigation. The core of the thesis lies...
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Function of alternative sigma factors of RNA polymerase in stress response of Corynebacterium glutamicum
Dostálová, Hana ; Nešvera, Jan (advisor) ; Lichá, Irena (referee)
The aim of this thesis was to contribute to understanding of the functions of the alternative sigma factors of RNA polymerase, σH a σM , during stress response of C. glutamicum. The role of σH and σM in the transcription of the gene sigM encoding σM and the genes of the operon dnaK-grpE-dnaJ-hspR which encode proteins involved in heat-shock response of C. glutamicum was studied. The promoters of the tested genes were cloned into the "promoter-probe" vector pET2 and their activity was determined by the measuring of specific activity of the reporter enzyme chloramfenicol acetyltransferase. It was found, that the heat stress has a moderate positive effect on the activity of the promoter of the gene sigM (P-sigM), whereas no effect of the oxidative stress induced by diamide was found. It was proved, that deletion of the gene sigH or sigM itself does not lead to decrease of activity of the promoter P-sigM neither in standard conditions nor after heat-shock. On the other hand, complete abolition of the activity of the promoter P-sigM was observed in the strain C. glutamicum ∆sigH∆sigM. The promoter of the gene sigM is thus recognized in standard conditions and after heat stress is recognized by both sigma factors σH and σM , but not by the sigma factor σA . It was shown, that mutation in the -10 region...
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Statistical analysis of the airline transport by appropriate quantitative methods
Dostálová, Hana ; Petera, Jiří (advisor) ; Lenka, Lenka (referee)
The theme of the air transport is becoming increasingly popular among people nowadays. Therefore, this Master thesis deals with a statistical analysis of the Czech airline Czech Airlines j.s.c. The system of timelines from 2003 to 2014 was used to analyse the number of transported passengers regarding passenger traffic and the number of transported goods and mail within the air freight transport.
The theoretical part of the paper is based on a literary review where basic terms of the air transport and timelines are described. The aim of the analysis is to apply particular trendy features on indicators of the research. According to the selected trend, predictions for the following the three seasons are made.
Subsequently, the statistical analysis is evaluated and recommendations aimed
at improving the future situation of the company are made.
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