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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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Nuclear lamina, its function in the cell and its role in interaction with non-enveloped DNA viruses
Pápež, Samuel ; Bruštíková, Kateřina (advisor) ; Kuthan, Martin (referee)
The nuclear lamina is a protein network found in all metazoa. It is composed of intermediate filaments V, namely type A and type B lamins. It is a multifunctional cellular structure adjacent to the inner nuclear membrane, giving the nucleus its shape, ensuring its mechanical stability and participating in the regulation of many nuclear processes. The nuclear lamina also has a protective function manifested in the infection of cells with DNA viruses, whose life cycle takes place in the nucleoplasm. These include small non-enveloped DNA viruses. The nuclear lamina is an obstacle that viruses have had to learn to overcome in order to multiply. To do this, they use their own, but also cellular mechanisms, which disrupt the nuclear membrane together with the nuclear lamina and thus allow the virus nuclear entry and exit. This work summarizes the knowledge about changes in the nuclear lamina induced by viruses of the families Polyomaviridae, Parvoviridae and Circoviridae, on the way through the nuclear envelope. Keywords: Nuclear lamina, disassembly of nuclear lamina, DNA viruses, Polyomaviruses, Parvoviruses, Circoviruses
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