Mutations in the putative calcium-binding domain of polyomavirus VP1 affect capsid assembly

Haynes, John I.; Chang, Deching; Consigli, Richard A.

doi:10.1128/jvi.67.5.2486-2495.1993

Cited by 43 publications

(26 citation statements)

References 48 publications

(56 reference statements)

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“…Though EF-hand-containing proteins have been found abundantly in eukaryotes and bacteria, the literature on EF-hand or EF-hand like Ca 2+ -binding motifs in virus proteins is thin, possibly due to lack of accurate prediction methods and robust methodologies for validation. A thorough search in PubMed with the key words "EF-hand and virus" only resulted in four hits of predicted canonical EF-hand motifs: the transmembrane protein gp 41 of HIV-1 [104], VP1 of polyomavirus [129], the VP7 outer capsid protein of rotavirus [102], and the protease domain of rubella virus (discussed below) [130]. Recombinant VP1 of an avian polyomavirus that lacks the 12residue predicted EF-hand loop is unable to bind 45 Ca and fails to assemble capsomeres into capsid-like particles [129].…”

Section: Continuous Viral Ca 2+ -Binding Sitesmentioning

confidence: 99%

“…A thorough search in PubMed with the key words "EF-hand and virus" only resulted in four hits of predicted canonical EF-hand motifs: the transmembrane protein gp 41 of HIV-1 [104], VP1 of polyomavirus [129], the VP7 outer capsid protein of rotavirus [102], and the protease domain of rubella virus (discussed below) [130]. Recombinant VP1 of an avian polyomavirus that lacks the 12residue predicted EF-hand loop is unable to bind 45 Ca and fails to assemble capsomeres into capsid-like particles [129]. However, the structural and Ca 2+ -binding properties of the first three reported putative canonical EF-hands have not been characterized yet.…”

Section: Continuous Viral Ca 2+ -Binding Sitesmentioning

confidence: 99%

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Viral calciomics: Interplays between Ca2+ and virus

2009

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Ca2+ is one of the most universal and versatile signaling molecules and is involved in almost every aspect of cellular processes. Viruses are adept at utilizing the universal Ca2+ signal to create a tailored cellular environment that meets their own demands. This review summarizes most of the known mechanisms by which viruses perturb Ca2+ homeostasis and utilize Ca2+ and cellular Ca2+-binding proteins to their benefit in their replication cycles. Ca2+ plays important roles in virion structure formation, virus entry, viral gene expression, posttranslational processing of viral proteins and virion maturation and release. As part of the review, we introduce an algorithm to identify linear “EF-hand” Ca2+-binding motifs which resulted in the prediction of a total of 93 previously unrecognized Ca2+-binding motifs in virus proteins. Many of these proteins are nonstructural proteins, a class of proteins among which Ca2+ interactions had not been formerly appreciated. The presence of linear Ca2+-binding motifs in viral proteins enlarges the spectrum of Ca2+–virus interplay and expands the total scenario of viral calciomics.

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Section: Continuous Viral Ca 2+ -Binding Sitesmentioning

confidence: 99%

Section: Continuous Viral Ca 2+ -Binding Sitesmentioning

confidence: 99%

Viral calciomics: Interplays between Ca2+ and virus

2009

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“…However, both mutant proteins were defective in self-assembly into VLPs. Although VP1 Ala pentamers looked indistinguishable from those of VP1 wt , they were not able (in contrast to the wild-type ones) to form VLPs after addition of CaCl 2 [11].…”

Section: Introductionmentioning

confidence: 76%

“…VP1 also binds Ca 2+ ions and the calcium-binding domain is localised between residues 266 and 277 [11]. This amino acid stretch is conserved in Polyomaviridae, suggesting an importance of calcium for PyV virions.…”

Section: Introductionmentioning

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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“…Either a reducing agent or a metal ion chelator alone can not disrupt the capsid particle [17,19]. Instead, capsomeres can be reassembled into a capsid‐like structure in the presence of calcium ions alone without the involvement of disulfide bonds [18,20,21]. Therefore, the role of disulfide bonds in polyomavirus assembly needs to be further clarified.…”

Section: Introductionmentioning

confidence: 99%

Disulfide bonds stabilize JC virus capsid‐like structure by protecting calcium ions from chelation

Chen

Wang

et al. 2001

FEBS Letters

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To investigate the role of disulfide bonds in the capsid structure, a recombinant JC virus-like particle (VLP) was used. The major capsid protein, VP1, of the JC virus was expressed in yeast cells. The yeast-expressed VP1 was self-assembled into a VLP. Disulfide bonds were found in the VLP which caused dimeric and trimeric VP1 linkages as demonstrated by nonreducing SDS^PAGE. The VLP remained intact when disulfide bonds were reduced by dithiothreitol. The VLP without disulfide bonds could be disassembled into capsomeres by EGTA alone, but those with disulfide bonds could not be disassembled by EGTA. Capsomeres were reassembled into VLPs in the presence of calcium ions. Capsomeres formed irregular aggregations instead of VLPs when treated with diamide to reconstitute the disulfide bonds. These results indicate that disulfide bonds play an important role in maintaining the integrity of the JC VLP by protecting calcium ions from chelation. ß 2001 Published by Elsevier Science B.V. on behalf of the Federation of European Biochemical Societies.

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Mutations in the putative calcium-binding domain of polyomavirus VP1 affect capsid assembly

Abstract: Calcium ions appear to play a major role in maintaining the structural integrity of the polyomavirus and are likely involved in the processes of viral uncoating and assembly. Previous studies demonstrated that a VP1

Cited by 43 publications

References 48 publications

Viral calciomics: Interplays between Ca2+ and virus

Viral calciomics: Interplays between Ca2+ and virus

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

Disulfide bonds stabilize JC virus capsid‐like structure by protecting calcium ions from chelation

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