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Role of nuclear lamins and Nup358 in BK polyomavirus infection
Išler, Lukáš ; Bruštíková, Kateřina (advisor) ; Němečková, Šárka (referee)
BK polyomavirus (BKPyV) has been causing serious health complication for several decades, especially in immunocompromised patients. The aim of this work was to investigate the progress of BKPyV replication during infection of human cells, as well as the influence of BKPyV infection on the nuclear lamina and nuclear pore proteins (NPC; Nuclear Pore Complex). During characterization and comparison of BKPyV infection in RPTEC/hTERT1 and MRC-5 cells we showed that BKPyV replicates better in RPTEC/hTERT1 cells as the productive infection results in six times higher viral titer. Using confocal microscopy we did not observe any nuclear lamina disruption nor VP1 accumulation under nuclear lamina that was previuosly observed in mouse polyomavirus infection. We verified previous observations of cytoplasmic deposits of NPC colocalizing with VP1 protein 24 hours post infection (hpi) with BKPyV and we showed that Nup358, protein of NPC, is a component of NPC deposits colocalizing with VP1. Neither transient expression from vectors encoding late region of BKPyV genome (pEF- BKV-late) or VP1 alone (pIaW), nor LT antigen expression analysis did not suggest any conection of observed phenomena to the productive infection. However, pseudoinfection of RPTEC/hTERT1 cells with VLPs derived from BKPyV induced VP1...
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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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