Cooperation of structural proteins during late events in the life cycle of polyomavirus

Forstová, Jitka; Krauzewicz, Nina; Wallace, Susan S.; Street, Alasdair J.; Dilworth, S. J.; Beard, Stuart; Griffin, Beverly E.

doi:10.1128/jvi.67.3.1405-1413.1993

Cited by 112 publications

(74 citation statements)

References 41 publications

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“…The proteins with EGFP in the opposite orientation (EGFP-VP2, EGFP-VP3) were targeted preferentially into the cell nucleus, and had markedly lower affinity to intracellular membranes. In agreement with our results, previous studies have shown that minor proteins were not targeted efficiently into the nucleus in insect cells or in African Green monkey kidney cells when VP1 was not co-expressed [8,9].…”

Section: Discussionsupporting

confidence: 93%

“…During MPyV infection, newly synthesized structural proteins form complexes (5VP1-1VP2, or 5VP1-1VP3) in the cytoplasm, which are then transported into the cell nucleus [8,9,19]. In the absence of VP1, we found that a substantial amount of VP2 or VP3 in the cytoplasm was co-localized with intracellular membranes, similar to the observation with fusion variants where EGFP was connected to their C-termini (VP2-EGFP, VP3-EGFP).…”

Section: Discussionsupporting

confidence: 83%

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Minor capsid proteins of mouse polyomavirus are inducers of apoptosis when produced individually but are only moderate contributors to cell death during the late phase of viral infection

et al. 2010

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Minor structural proteins of mouse polyomavirus (MPyV) are essential for virus infection. To study their properties and possible contributions to cell death induction, fusion variants of these proteins, created by linking enhanced green fluorescent protein (EGFP) to their C-or N-termini, were prepared and tested in the absence of other MPyV gene products, namely the tumor antigens and the major capsid protein, VP1. The minor proteins linked to EGFP at their C-terminus (VP2-EGFP, VP3-EGFP) were found to display properties similar to their nonfused, wild-type versions: they killed mouse 3T3 cells quickly when expressed individually. Carrying nuclear localization signals at their common C-terminus, the minor capsid proteins were detected in the nucleus. However, a substantial subpopulation of both VP2 and VP3 proteins, as well as of the fusion proteins VP2-EGFP and VP3-EGFP, was detected in the cytoplasm, co-localizing with intracellular membranes. Truncated VP3 protein, composed of 103 C-terminal amino acids, exhibited reduced affinity for intracellular membranes and cytotoxicity. Biochemical studies proved each of the minor proteins to be a very potent inducer of apoptosis, which was dependent on caspase activation. Immunoelectron microscopy showed the minor proteins to be associated with damaged membranes of the endoplasmic reticulum, nuclear envelope and mitochondria as soon as 5 h post-transfection. Analysis of apoptotic markers and cell death kinetics in cells transfected with the wild-type MPyV genome and the genome mutated in both VP2 and VP3 translation start codons revealed that the minor proteins contribute moderately to apoptotic processes in the late phase of infection and both are dispensable for cell destruction at the end of the virus replication cycle. Structured digital abstractl MINT-7386399, MINT-7386463, MINT-7386515: VP3 (uniprotkb:P03096-2) and GRP94

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Section: Discussionsupporting

confidence: 93%

Section: Discussionsupporting

confidence: 83%

Minor capsid proteins of mouse polyomavirus are inducers of apoptosis when produced individually but are only moderate contributors to cell death during the late phase of viral infection

et al. 2010

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“…Cells were harvested 72 h post infection (p.i. ), lysed, and VLPs were purified by CsCl and sucrose gradients as described previously [8].…”

Section: Isolation Of Capsid-like Particles From Insect Cellsmentioning

confidence: 99%

Polyomavirus EGFP‐pseudocapsids: Analysis of model particles for introduction of proteins and peptides into mammalian cells

et al. 2005

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A vector for preparation of mouse polyomavirus capsid-like particles for transfer of foreign peptides or proteins into cells was constructed. Model pseudocapsids carrying EGFP fused with the C-terminal part of the VP3 minor protein (EGFP-VLPs) have been prepared and analysed for their ability to be internalised and processed by mouse cells and to activate mouse and human dendritic cells (DC) in vitro. EGFP-VLPs entered mouse epithelial cells, fibroblasts and human and mouse DC efficiently and were processed by both, lysosomes and proteasomes. Surprisingly, they did not induce upregulation of DC co-stimulation molecules or maturation markers in vitro; however, they did induce interleukin 12 secretion.

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“…The recombinant plasmid p425GVP1 wt (kindly provided by Mgr. L. Synek (Charles University, Prague, Czech Republic)) and p426GVP1 wt was constructed by ligation of a BamHI-BglII fragment containing the VP1 gene cut from pVL1393-VP1 [13] and yeast shuttle expression vector p425G or p426G [24] cut with BamHI.…”

Section: Preparation Of Recombinant Plasmidsmentioning

confidence: 99%

Point mutation in calcium-binding domain of mouse polyomavirus VP1 protein does not prevent virus-like particle formation, but changes VP1 interactions with cell structures

Adamec

Palková

Velková

et al. 2005

FEMS Yeast Research

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The mouse polyomavirus gene for the major structural protein, VP1, with point mutation in the calcium-binding pocket (VP1(Ala)), was expressed in Saccharomyces cerevisiae and in a baculovirus expression system. Surprisingly, VP1(Ala) forms virus-like particles (VLPs) in nuclei of both yeast and insect cells. VP1(Ala)-VLPs produced in S. cerevisiae are unstable and, unlike wild-type VP1 (VP1(wt))-VLPs, they disassemble during the purification procedure and storage. In contrast to VP1(wt), VP1(Ala) does not interact with the yeast mitotic spindle. Nevertheless, both wild-type and mutated VP1 inhibit yeast cell growth. The inhibition is cAMP-dependent. The production of VP1(Ala) and VP1(wt)-VLPs in insect cells also revealed differences in their interactions with cellular protein(s). Thus, the mutation in the VP1 calcium pocket alters the stability and surface conformation of VLPs rather than the ability of VP1 to self-assemble.

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Cooperation of structural proteins during late events in the life cycle of polyomavirus

Cited by 112 publications

References 41 publications

Minor capsid proteins of mouse polyomavirus are inducers of apoptosis when produced individually but are only moderate contributors to cell death during the late phase of viral infection

Minor capsid proteins of mouse polyomavirus are inducers of apoptosis when produced individually but are only moderate contributors to cell death during the late phase of viral infection

Polyomavirus EGFP‐pseudocapsids: Analysis of model particles for introduction of proteins and peptides into mammalian cells

Point mutation in calcium-binding domain of mouse polyomavirus VP1 protein does not prevent virus-like particle formation, but changes VP1 interactions with cell structures

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