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Interactions of polyomavirus structures with components of cell innate immunity
Portychová, Tereza ; Forstová, Jitka (advisor) ; Schreiberová, Lucie (referee)
The topic of this thesis are the interactions of polyomavirus structures with components of innate immunity in infected cells. This review is focused on model SV40 and MPyV polyomaviruses and human BKPyV, JCPyV and MCPyV. The research of the interplay of innate immunity response and polyomaviruses is in its infancy. Infection with all studied polyomaviruses induces, via their LT antigens, DNA damage response (DDR), necessary for their efficient replication. DDR can activate both the canonical and the non-canonical pathway of interferon induction leading to an antiviral state. Polyomaviruses are recognized by the immune system first during replication of their genomes. Interferon induction by polyomaviruses can be initiated by the DNA sensor, cGAS, followed by STING activation, but also by recognition of the viral RNA by the RIG-1 sensor. The virus early LT and st antigens and the late agnoprotein of some polyomaviruses have demonstrated the potential to regulate innate immune responses and thus contribute to the establishment of polyomavirus persistence. Keywords: polyomaviruses, innate cell immunity, large T antigen, small t antigen, agnoprotein, interferon-stimulated genes
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Experimental system for the mouse polyomavirus life cycle study
Pergner, Jiří ; Španielová, Hana (advisor) ; Mašek, Tomáš (referee)
Experimental system for the mouse polyomavirus life cycle study Abstract: Murine polyomavirus (MPyV) is the prototype of the Polyomaviridae family. This family includes also some important human pathogens (BKV, JCV, Merkel cell polyomavirus). Due to their specific properties viruses within this family may serve as versatile vectors for gene therapy or recombinant vaccine production. New methodological approaches may help to understand some yet unknown facts about MPyV life cycle. Clarification of some processes during murine polyomavirus life cycle may be also important to fully exploit polyomaviruses for therapeutic purposes. The aim of this diploma thesis was to preparare two innovative experimental systems that extend possibilities of studying the life cycle of MPyV. The first part of the diploma thesis focusses on construction of recombinant MPyV which expresses yellow fluorescent protein (EYFP) in the early stages of infection. Such virus can be very useful for studying the infection spreading by live- cell imaging and Fluorescence-Activated Cell Sorting (FACS) and can be employed for co- localization studies of YFP-tagged LT antigen with certain cellular proteins. Second part of the diploma thesis describes preparation of a hybrid cell line prepared by fusion of mouse and monkey cells. This new cell...
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The noncoding control region of human polyomaviruses
Pešek, David ; Saláková, Martina (advisor) ; Váňová, Jana (referee)
Genome of human polyomavirus consists of circular dsDNA around 5000 base and can be divided into three functional regions - the early viral gene region (EVGR), that encodes the regulatory T antigen and miRNAs, noncoding control region (NCCR) harboring the minimal cis- acting elements involved in viral replication and the late viral gene region (LVGR), that encodes the structural capsid proteins. Noncoding control region contains the origin of viral replication that overlaps the promoters that control expresion of early and late gene region. Noncoding control region sequences include a large number of various binding sites for cellular transcription factors involved in regulation expression from LVGR and EVGR. This thesis describes the organization of the most variable region of the PyV genome, NCCR, in chosen polyomaviruses SV40, BKPyV and JCPyV. This region often undergoes rearrangements, deletion and point mutations that affects exression of human polyomavirus. Key words: polyomavirus, noncoding control region, BKPyV virus, JCPyV virus, SV40, large T antigen, transcriptional factor
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Experimental system for the mouse polyomavirus life cycle study
Pergner, Jiří ; Mašek, Tomáš (referee) ; Španielová, Hana (advisor)
Experimental system for the mouse polyomavirus life cycle study Abstract: Murine polyomavirus (MPyV) is the prototype of the Polyomaviridae family. This family includes also some important human pathogens (BKV, JCV, Merkel cell polyomavirus). Due to their specific properties viruses within this family may serve as versatile vectors for gene therapy or recombinant vaccine production. New methodological approaches may help to understand some yet unknown facts about MPyV life cycle. Clarification of some processes during murine polyomavirus life cycle may be also important to fully exploit polyomaviruses for therapeutic purposes. The aim of this diploma thesis was to preparare two innovative experimental systems that extend possibilities of studying the life cycle of MPyV. The first part of the diploma thesis focusses on construction of recombinant MPyV which expresses yellow fluorescent protein (EYFP) in the early stages of infection. Such virus can be very useful for studying the infection spreading by live- cell imaging and Fluorescence-Activated Cell Sorting (FACS) and can be employed for co- localization studies of YFP-tagged LT antigen with certain cellular proteins. Second part of the diploma thesis describes preparation of a hybrid cell line prepared by fusion of mouse and monkey cells. This new cell...
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