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Cloning, expression and characterization of the viral proteins
Pekárek, Matěj ; Šilhán, Jan (advisor) ; Obšil, Tomáš (referee)
Over the last twenty years three new coronavirus strains have appeared in human population, that cause dangerous infections of respiratory tract. At this moment no approved drug exists, that could mitigate the infection caused by these viruses. One of the possible targets of these drugs are proteins responsible for viral genome replication. Nsp13 is primarily a helicase, that unwinds double stranded nucleic acids in the direction of 5'-3', but it also increases activity of replication and transcription complex, it participates in the synthesis of 5' cap on viral RNA and it probably mediates the backtracking of replication and transcripton complex during the process of RNA repair. On top of that nsp13 is strongly conserved among all coronaviruses and it is possible target not only for COVID-19 treatment, but also infections caused by potential future coronaviruses. Theoretical part of the thesis briefly describes the life cycle of SARS-CoV-2, functions of individual proteins of this virus, that participate in replication and transcription of RNA and it summarizes current knowledge about function of nsp13 and its potential inhibitors. In experimental part we have prepared the expression vector for nsp13, that was used for the production of nsp13. Nsp13 was then isolated and purified. (In Czech) Key...
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Host-virus interactions of mammalian endogenous retroviruses
Farkašová, Helena ; Elleder, Daniel (advisor) ; Hirsch, Ivan (referee) ; Mělková, Zora (referee)
Endogenous retroviruses (ERVs) originate by germline infection and subsequent mendelian inheritance of their exogenous counterparts. With notable exceptions, all mammalian ERVs are evolutionarily old and fixed in the population of its host species. Some groups of retroviruses were believed not to be able to form endogenous copies. We discovered an additional endogenous Lentivirus and a first endogenous Deltaretrovirus. Both of these groups were previously considered unable to form endogenous copies. Endogenous lentiviruses were discovered only recently and are still quite rare. These are still just small pieces of evidence insufficient to give a broader picture about the history of virus endogenization. We described a novel endogenous Lentivirus in the genome of Malayan colugo (Galeopterus variegatus) denoted ELVgv (endogenous Lentivirus of G. variegatus). Based on several analyses we proved that this is the oldest Lentivirus discovered up to date and confirmed its presence in the only other extant species of Dermoptera - Cynocephalus volans. Endogenous deltaretroviruses were the last group without a single endogenous member. We detected the remnants of endogenous Deltaretrovirus in the genome of Natal Long-fingered bat (Miniopterus natalensis). However, this sequence was present in the genome only in one...
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Implication of eukaryotic DNA repair machinery in viral replication
Hron, Tomáš ; Španielová, Hana (advisor) ; Harant, Karel (referee)
Eukaryotic DNA damage response is an important mechanism which ensures genome stability. Its components are also mobilized during viral infection as a reaction against extraneous nucleic acid. Additionally, DNA repair machinery seems to be activated by some viruses purposely to provide their replication. This activation is mediated mainly by viral proteins which are able to interact with cellular factors. In many cases, key components of DNA damage mechanisms are associated with viral replication centre and likely participate in this process. Furthermore, cellular DNA damage signaling is exploited to provide competent environment for viral reproduction. However, particular mechanisms how these cellular factors participate in viral infection are still largely unclear. In this thesis, the principles of relationship between viral infection and eukaryotic DNA damage response are summarized and main viral families which are known to activate and utilize these mechanisms for its genom replication are described.
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Host-virus interactions of mammalian endogenous retroviruses
Farkašová, Helena
Endogenous retroviruses (ERVs) originate by germline infection and subsequent mendelian inheritance of their exogenous counterparts. With notable exceptions, all mammalian ERVs are evolutionarily old and fixed in the population of its host species. Some groups of retroviruses were believed not to be able to form endogenous copies. We discovered an additional endogenous Lentivirus and a first endogenous Deltaretrovirus. Both of these groups were previously considered unable to form endogenous copies. Endogenous lentiviruses were discovered only recently and are still quite rare. These are still just small pieces of evidence insufficient to give a broader picture about the history of virus endogenization. We described a novel endogenous Lentivirus in the genome of Malayan colugo (Galeopterus variegatus) denoted ELVgv (endogenous Lentivirus of G. variegatus). Based on several analyses we proved that this is the oldest Lentivirus discovered up to date and confirmed its presence in the only other extant species of Dermoptera - Cynocephalus volans. Endogenous deltaretroviruses were the last group without a single endogenous member. We detected the remnants of endogenous Deltaretrovirus in the genome of Natal Long-fingered bat (Miniopterus natalensis). However, this sequence was present in the genome only in one...
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Host-virus interactions of mammalian endogenous retroviruses
Farkašová, Helena
Endogenous retroviruses (ERVs) originate by germline infection and subsequent mendelian inheritance of their exogenous counterparts. With notable exceptions, all mammalian ERVs are evolutionarily old and fixed in the population of its host species. Some groups of retroviruses were believed not to be able to form endogenous copies. We discovered an additional endogenous Lentivirus and a first endogenous Deltaretrovirus. Both of these groups were previously considered unable to form endogenous copies. Endogenous lentiviruses were discovered only recently and are still quite rare. These are still just small pieces of evidence insufficient to give a broader picture about the history of virus endogenization. We described a novel endogenous Lentivirus in the genome of Malayan colugo (Galeopterus variegatus) denoted ELVgv (endogenous Lentivirus of G. variegatus). Based on several analyses we proved that this is the oldest Lentivirus discovered up to date and confirmed its presence in the only other extant species of Dermoptera - Cynocephalus volans. Endogenous deltaretroviruses were the last group without a single endogenous member. We detected the remnants of endogenous Deltaretrovirus in the genome of Natal Long-fingered bat (Miniopterus natalensis). However, this sequence was present in the genome only in one...
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Host-virus interactions of mammalian endogenous retroviruses
Farkašová, Helena ; Elleder, Daniel (advisor) ; Hirsch, Ivan (referee) ; Mělková, Zora (referee)
Endogenous retroviruses (ERVs) originate by germline infection and subsequent mendelian inheritance of their exogenous counterparts. With notable exceptions, all mammalian ERVs are evolutionarily old and fixed in the population of its host species. Some groups of retroviruses were believed not to be able to form endogenous copies. We discovered an additional endogenous Lentivirus and a first endogenous Deltaretrovirus. Both of these groups were previously considered unable to form endogenous copies. Endogenous lentiviruses were discovered only recently and are still quite rare. These are still just small pieces of evidence insufficient to give a broader picture about the history of virus endogenization. We described a novel endogenous Lentivirus in the genome of Malayan colugo (Galeopterus variegatus) denoted ELVgv (endogenous Lentivirus of G. variegatus). Based on several analyses we proved that this is the oldest Lentivirus discovered up to date and confirmed its presence in the only other extant species of Dermoptera - Cynocephalus volans. Endogenous deltaretroviruses were the last group without a single endogenous member. We detected the remnants of endogenous Deltaretrovirus in the genome of Natal Long-fingered bat (Miniopterus natalensis). However, this sequence was present in the genome only in one...
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Implication of eukaryotic DNA repair machinery in viral replication
Hron, Tomáš ; Španielová, Hana (advisor) ; Harant, Karel (referee)
Eukaryotic DNA damage response is an important mechanism which ensures genome stability. Its components are also mobilized during viral infection as a reaction against extraneous nucleic acid. Additionally, DNA repair machinery seems to be activated by some viruses purposely to provide their replication. This activation is mediated mainly by viral proteins which are able to interact with cellular factors. In many cases, key components of DNA damage mechanisms are associated with viral replication centre and likely participate in this process. Furthermore, cellular DNA damage signaling is exploited to provide competent environment for viral reproduction. However, particular mechanisms how these cellular factors participate in viral infection are still largely unclear. In this thesis, the principles of relationship between viral infection and eukaryotic DNA damage response are summarized and main viral families which are known to activate and utilize these mechanisms for its genom replication are described.
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