National Repository of Grey Literature 78 records found  1 - 10nextend  jump to record: Search took 0.00 seconds. 
Searching for interaction partners associated with the transcriptional apparatus of yeast linear plasmids
Ľalíková, Kristýna ; Vopálenský, Václav (advisor) ; Čáp, Michal (referee)
The pGKL plasmids are a type of yeast linear double-stranded DNA plasmids found in the cytoplasm of yeast Kluyveromyces lactis. The plasmid gene transcription process involves the plasmid transcription apparatus, which shows similarity to the transcription apparatus of poxviruses. Poxviruses cause a range of serious diseases. Thus pGKL plasmids offer a safe way to study some of the mechanisms of these viruses without the risk of infection. The plasmid transcription apparatus comprises non-canonical RNA polymerase, helicase, and capping enzyme as its main components. Although RNA polymerase has been thoroughly characterized and described, little focus has been on the interactions between proteins within the transcription apparatus. Therefore, this study aims to shed light on this fundamental aspect of the gene expression of pGKL plasmids. The main goal of the thesis was to prepare a system that would facilitate the search for interaction partners of the pGKL transcription apparatus and verify the interactions between chosen proteins in yeast cells. The foundation of this system relies on the use of optimized genes and the yeast two-hybrid system method. The development of the experimental method has begun successfully, and it's expected to conclude soon, although further optimizations are still...
Post-transcriptional modification of mRNA molecules in viruses of the Poxviridae family
Jakešová, Kristýna ; Vopálenský, Václav (advisor) ; Šmahel, Michal (referee)
Post-transcriptional modifications of mRNA molecules in viruses belonging to the Poxviridae family, including the vaccinia virus, play a key role in regulating viral gene expression and also influence virus-host cell interactions. These modifications include the synthesis of a 7-methylguanosine cap, 2'-O-methylation, 3' polyadenylation of the mRNA strand, hydrolysis of the 7-methylguanosine cap, and 5' polyadenylation of the mRNA strand. Studying post-transcriptional or co-transcriptional mRNA modifications in Poxviridae viruses can contribute to a better understanding of viral pathogenesis and the development of new strategies for treating infections caused by viruses in this family. The aim of this bachelor's thesis is to summarize current knowledge of co-transcriptional or post-transcriptional mRNA modifications in Poxviridae viruses, with a special focus on vaccinia virus.
Comparative analysis of RNA binding properties of Rpl22 proteins
Gredová, Alexandra ; Abrhámová, Kateřina (advisor) ; Vopálenský, Václav (referee)
After the whole genome duplication event, Saccharomyces cerevisiae lost 90 % of its paralogs. 59 ribosomal protein genes (RPG) retained a duplicated form. The cell has to balance expression of RPGs as a part of adaptation to changing conditions, thus ensuring the production of the right ratio of ribosomal proteins (RP). In addition, RPGs contain 1/3 of all introns found in the S. cerevisiae genome. Rpl22A/B are part of the large ribosomal subunit where they contact 25S rRNA. Deletion of these RPGs results in 2X slower growth in comparison with WT cells. Within their extraribosomal function Rpl22A/B are able to intragenically and intergenically regulate their expression. Binding of Rpl22A/B to the intronic part of pre-mRNA of RPL22A or RPL22B results in splicing inhibition, which is stronger in the case of the RPL22B intron (RPL22Bi). However, the exact mechanism is not known. We know that Rpl22s from various organisms bind a hairpin structure that can be found in 25S rRNA also. Since the predicted structure of RPL22Bi does not contain this binding motif, it rises a question of whether the RNA binding surface of Rpl22A/B is different or more extensive than the one with which Rpl22 contacts 25S rRNA. We compared the ability of Rpl22 from different organisms to complement the functions of Rpl22A/B. These...
Non-template activities of DNA/RNA polymerases and their significance
Pokorná, Kristýna ; Vopálenský, Václav (advisor) ; Pospíšil, Jiří (referee)
Non-templated addition of adenosine on the 3' end of the product of several DNA polymerases commonly used in molecular biology has been a problematic yet unexplained phenomenon for a few decades now. The only other instance where adenosine is added to the 3' end of an information molecule in a cell is polyadenylation. At the same time, several RNA viruses appear to perform similar action to either edit or polyadenylate their mRNAs. This work focuses on the details of these three highlighted processes and relationship between them and has found out that all of them appear to stem from a common conserved property of the ancestral polymerase. Key words: DNA polymerase, RNA polymerase, non-templated addition, Taq polymerase, polyadenylation
Effect of ADAR1 enzyme on hepatitis C virus life cycle
Kubů, Martin ; Vopálenský, Václav (advisor) ; Fraiberk, Martin (referee)
Hepatitis C virus (HCV) is a virus of the family Flaviviridae whose genome consists of ,,+RNA" molecule. It causes the disease hepatitis Cepatitida typu C, which affects tens of millions of people worldwide. Although there is an effective treatment for this type of hepatitis, a preventive vaccine against the HCV virus has not yet been developed. Several ambigious works focused on the relationship between HCV and the enzyme adenosine deaminase acting on double-stranded RNA 1 (ADAR1) were published in the past. This dimeric double-stranded RNA binding enzyme is a part of innate immunity and causes catalytic conversion of adenosine to inosine, which is recognized by cellular mechanisms as guanine, which ultimately leads to mutations in the affected dsRNA molecule. The works published so far attribute an antiviral function to the ADAR1 enzyme in the context of HCV infection. However, vectors containing the entire HCV genome were not used in these works, and a cell line with deletion od the ADAR1 gene has never been used so far. The actual relationship between the ADAR1 enzyme and the HCV virus has not yet been reliably verified. The aim of this thesis was to gain a deeper understanding of the relationship between the HCV virus and the ADAR1 enzyme. For this task, HCV permissive Huh7.5 cell lines with...
delta subunit of bacterial RNA pol and its role in regulation of gene expression in B. subtilis
Dvořáček, Lukáš ; Krásný, Libor (advisor) ; Vopálenský, Václav (referee)
Delta subunit of bacterial RNA pol and its role in regulation of gene expression in B. subtilis. In this work I focus on regulation of eubacterial gene expression. First, I describe recent knowledge about a key stage of gene expression - transcription, focusing on regulation of trancription iniciation via small effector molecules (guanosine tetraphosphate, initiating nucleoside triphosphate) that are important for the regulation of ribosomal RNA. Second, in the experimental part of my work, I focus on the role of the _ protein, a subunit of RNA polymarase in gram positive bacteria, in transcription iniciation and its effects on regulation of RNA polymerase by the concentration of initiating nucleoside triphosphates.

National Repository of Grey Literature : 78 records found   1 - 10nextend  jump to record:
Interested in being notified about new results for this query?
Subscribe to the RSS feed.