Mutations of polyomavirus VP1 allow in vitro growth in undifferentiated cells and modify in vivo tissue replication specificity

Mezes, Beata; Amati, Paolo

doi:10.1128/jvi.68.2.1196-1199.1994

Cited by 12 publications

(4 citation statements)

References 31 publications

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“…A determinant responsible for efficient virus spread and induction of tumors has been mapped to a single amino acid in the major capsid protein VP1 (9,13). Determinants that alter the spectrum of tissues in which polyomavirus can replicate in the mouse have also been mapped to VP1 (22). Mutations in the tumor antigens that block or reduce the virus's ability to transform cells in vitro (18,23) inhibit or alter its tumorigenicity in the mouse (9,11,14,30).…”

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confidence: 99%

Genetic and structural analysis of a virulence determinant in polyomavirus VP1

Bauer

Bronson

Fung

et al. 1995

J Virol

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The LID strain of polyomavirus differs from other laboratory strains in causing a rapidly lethal infection of newborn C3H/Bi mice. This virulent behavior of LID was attenuated by dilution, yet at sublethal doses LID was able to induce tumors at a high frequency, like its parent virus PTA. By constructing and assaying LID-PTA recombinant viruses and by DNA sequencing, the determinant of virulence in LID was mapped to the major viral capsid protein, VP1. The VP1s of LID and PTA differed at two positions: at 185, LID has phenylalanine and PTA has tyrosine, and at 296, LID has alanine and PTA has valine. Results obtained with viruses constructed by site-directed mutagenesis showed that alanine at position 296 is sufficient to confer a fully virulent phenotype regardless of which amino acid is at position 185. However, with valine at position 296, an effect of phenylalanine at position 185 is apparent, as this virus possesses an intermediate level of virulence. A crystal structure of polyomavirus complexed with 3-sialyl lactose previously indicated van der Waals contacts between the side chain of valine 296 and the sialic acid ring (T. Stehle, Y. Yan, T. L. Benjamin, and S. C. Harrison, Nature [London] 369:160-163, 1994). When this interaction was modeled with alanine, these contacts were greatly reduced. Direct confirmation that the substitutions in VP1 affected receptor binding was obtained by studying virus hemagglutination behavior. The ensemble of results are discussed in terms of the idea that a lower affinity of the virus for its receptor can result in more rapid spread and increased pathogenicity.

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confidence: 99%

Genetic and structural analysis of a virulence determinant in polyomavirus VP1

Bauer

Bronson

Fung

et al. 1995

J Virol

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“…VP1 1–136 shows to be totally unable to activate the reporter genes even in presence of YY1, therefore, indicating that aa 137–140 are required for VP1‐YY1 interaction in this experimental system and that the interacting sites with the yeast factor and YY1 are distinguishable. Since our previous results indicated that the D‐E loop of VP1 could be responsible for the viral‐host specificity [18,19] and may be involved in the formation of the complex of the viral genome with the NM (Amati et al, manuscript in preparation), we fused the sequences encoding the D‐E 118–170 or the E‐F 172–248 loops with the GAL4 DNA‐BD. The results of the 2HS as in Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Results from our and other laboratories [16–21] had suggested the possibility that VP1, the major capsid protein, could determine alterations in the host specificity of growth and tumorigenicity. In order to determine the possible mechanisms that allow the encapsidated polyomaviruses that reach the cell nucleus to associate with the NM, we investigated the possible role of YY1 in this necessary step of viral function.…”

Section: Introductionmentioning

confidence: 92%

The polyomavirus major capsid protein VP1 interacts with the nuclear matrix regulatory protein YY1

et al. 2000

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Polyomavirus reaches the nucleus in a still encapsidated form, and the viral genome is readily found in association with the nuclear matrix. This association is thought to be essential for viral replication. In order to identify the protein(s) involved in the virus-nuclear matrix interaction, we focused on the possible roles exerted by the multifunctional cellular nuclear matrix protein Yin Yang 1 (YY1) and by the viral major capsid protein VP1. In the present work we report on the in vivo association between YY1 and VP1. Using the yeast two-hybrid system we demonstrate that the VP1 and YY1 proteins physically interact through the D-E region of VP1 and the activation domain of YY1.z 2000 Federation of European Biochemical Societies.

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“…In addition, the phosphorylated Ser-63, Thr-66 and Thr-156 residues of murine polyomavirus VP1 are not conserved in the VP1 proteins of the other polyomaviruses. Thus, although these viruses belong to the same family, their VP1 sequences have low similarity and therefore modification of the capsid proteins may affect various biological functions, such as host range, tissue tropism and pathogenesis, in different ways (Freund et al, 1991;Mezes & Amati, 1994). In the current study, the phosphorylated residues of BK VP1, including Ser-80, Ser-133 and Ser-327, are distinct from those of polyomavirus VP1, where phosphorylation occurs at Ser-63, Thr-66 and Thr-156.…”

Section: Discussionmentioning

confidence: 99%

Phosphorylation of Ser-80 of VP1 and Ser-254 of VP2 is essential for human BK virus propagation in tissue culture

Chen

Hsu

Fang

et al. 2011

Journal of General Virology

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ROCBK virus (BKV) infection may cause polyomavirus-associated nephropathy in patients with renal transplantation. Recently, the phosphorylated amino acids on the structural proteins VP1, VP2 and VP3 of BKV have been identified by liquid chromatography-tandem mass spectrometry in our laboratory. In this study, we further analysed the biological effects of these phosphorylation events. Phosphorylation of the BKV structural proteins was demonstrated by [32 P]orthophosphate labelling in vivo. Site-directed mutagenesis was performed to replace all of the phosphorylated amino acids. The mutated BKV genomes were transfected into Vero cells for propagation analysis.The results showed that expression of the early protein LT and of the late protein VP1 by the mutants VP1-S80A, VP1-S80-133A, VP1-S80-327A, VP1-S80-133-327A and VP2-S254A was abolished. However, propagation of other mutants was similar to that of wild-type BKV. The results suggest that phosphorylation of Ser-80 of VP1 and Ser-254 of VP2 is crucial for BKV propagation. INTRODUCTIONPolyomaviruses are non-enveloped small DNA viruses and have a circular dsDNA genome of approximately 5.2 kbp. The polyomavirus family includes murine polyomavirus, simian virus (SV40), human John Cunningham virus (JCV), BK virus (BKV), KI virus, WU virus and Merkel cell polyomavirus (MCV) (Allander et al., 2007;Eash et al., 2006;Feng et al., 2008;Gardner et al., 1971;Gaynor et al., 2007;Kanitakis, 2008; Pérez-Losada et al., 2006). The genomes of these polyomaviruses contain regulatory, early and late regions. The regulatory region includes the replication origin as well as a promoter and an enhancer that regulate the transcription of the early and late genes. The early region encodes the regulatory proteins large tumour antigen (LT) and small tumour antigen, which are associated with replication and the regulation of the early and late genes. The late region encodes three structural capsid proteins, VP1, VP2 and VP3, and the regulatory agonoprotein (Safak et al., 2001). The viral capsid is composed of the major structural protein, VP1, the minor structural proteins, VP2 and VP3, and a viral minichromosome (Christiansen et al., 1977). The viral capsid consists of 72 pentameric capsomeres, including 12 pentavalent and 60 hexavalent capsomeres (Liddington et al., 1991).The structural proteins of polyomavirus play important roles in the infection life cycle, such as recognition of host receptors, haemagglutination activity (Bolen & Consigli, 1979;Bolen et al., 1981) and encapsidation of the viral genome during maturation (Chang et al., 1993;Gasparovic et al., 2006;Kawano et al., 2006). Furthermore, interactions among the structural proteins are also crucial for virus entry, nuclear translocation, DNA packaging and assembly of viral progeny (Chen et al., 1998;Delos et al., 1995;Gasparovic et al., 2006;Gharakhanian et al., 2003;Kawano et al., 2006;Shishido-Hara et al., 2004). However, it has been found that the structural proteins are modified into several subspecies, as demonstrated by t...

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Mutations of polyomavirus VP1 allow in vitro growth in undifferentiated cells and modify in vivo tissue replication specificity

Cited by 12 publications

References 31 publications

Genetic and structural analysis of a virulence determinant in polyomavirus VP1

Genetic and structural analysis of a virulence determinant in polyomavirus VP1

The polyomavirus major capsid protein VP1 interacts with the nuclear matrix regulatory protein YY1

Phosphorylation of Ser-80 of VP1 and Ser-254 of VP2 is essential for human BK virus propagation in tissue culture

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