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A structure-guided mutation in the major capsid protein retargets BK polyomavirus.

Neu U, Allen SA, Blaum BS, Liu Y, Frank M, Palma AS, Ströh LJ, Feizi T, Peters T, Atwood WJ, Stehle T - PLoS Pathog. (2013)

Bottom Line: We have characterized the receptor specificity, structure and infectivity of the human polyomavirus BKPyV, the causative agent of polyomavirus-associated nephropathy, and uncover a molecular switch for binding different carbohydrate receptors.The crystal structure of the BKPyV capsid protein VP1 in complex with GD3 reveals contacts with two sialic acid moieties in the receptor, providing a basis for the observed specificity.Mutation of this residue from lysine in BKPyV to serine in SV40 switches the receptor specificity of BKPyV from GD3 to GM1 both in vitro and in cell culture.

View Article: PubMed Central - PubMed

Affiliation: Interfaculty Institute of Biochemistry, University of Tübingen, Tübingen, Germany.

ABSTRACT
Viruses within a family often vary in their cellular tropism and pathogenicity. In many cases, these variations are due to viruses switching their specificity from one cell surface receptor to another. The structural requirements that underlie such receptor switching are not well understood especially for carbohydrate-binding viruses, as methods capable of structure-specificity studies are only relatively recently being developed for carbohydrates. We have characterized the receptor specificity, structure and infectivity of the human polyomavirus BKPyV, the causative agent of polyomavirus-associated nephropathy, and uncover a molecular switch for binding different carbohydrate receptors. We show that the b-series gangliosides GD3, GD2, GD1b and GT1b all can serve as receptors for BKPyV. The crystal structure of the BKPyV capsid protein VP1 in complex with GD3 reveals contacts with two sialic acid moieties in the receptor, providing a basis for the observed specificity. Comparison with the structure of simian virus 40 (SV40) VP1 bound to ganglioside GM1 identifies the amino acid at position 68 as a determinant of specificity. Mutation of this residue from lysine in BKPyV to serine in SV40 switches the receptor specificity of BKPyV from GD3 to GM1 both in vitro and in cell culture. Our findings highlight the plasticity of viral receptor binding sites and form a template to retarget viruses to different receptors and cell types.

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B-series gangliosides are receptors for BKPyV.(A) Schematic representation of structures of disialic acid-containing b-series gangliosides GD3, GD2, GD1b, GT1b and of the monosialylated a-series ganglioside GM1 indicating the abbreviated designations used here for individual residues in the gangliosides. (B) Ganglioside supplementation assays. Vero cells were incubated with gangliosides, challenged with BKPyV and scored for infection. The average number of VP1 positive cells is plotted compared to controls. Error bars represent the standard deviation for 3 independent experiments. Asterisks indicate p-value (*p<0.05). (C) STD off-resonance (top) and difference (bottom) spectra of WT BKPyV VP1 in the presence of 50-fold excess GD3 oligosaccharide. The off-resonance spectrum was scaled to 3%. GD3 resonances labeled in the difference spectrum receive considerable saturation transfer from the protein. Regions with strong signal overlap are not labeled because saturation effects in this region cannot be unambiguously assigned. Signals that were truncated are denoted by diagonal bars. (D) STD off-resonance (top) and difference spectrum (bottom) of wild type BKPyV in the presence of 50-fold excess GD1b oligosaccharide. The spectrum is labeled as in C.
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ppat-1003688-g001: B-series gangliosides are receptors for BKPyV.(A) Schematic representation of structures of disialic acid-containing b-series gangliosides GD3, GD2, GD1b, GT1b and of the monosialylated a-series ganglioside GM1 indicating the abbreviated designations used here for individual residues in the gangliosides. (B) Ganglioside supplementation assays. Vero cells were incubated with gangliosides, challenged with BKPyV and scored for infection. The average number of VP1 positive cells is plotted compared to controls. Error bars represent the standard deviation for 3 independent experiments. Asterisks indicate p-value (*p<0.05). (C) STD off-resonance (top) and difference (bottom) spectra of WT BKPyV VP1 in the presence of 50-fold excess GD3 oligosaccharide. The off-resonance spectrum was scaled to 3%. GD3 resonances labeled in the difference spectrum receive considerable saturation transfer from the protein. Regions with strong signal overlap are not labeled because saturation effects in this region cannot be unambiguously assigned. Signals that were truncated are denoted by diagonal bars. (D) STD off-resonance (top) and difference spectrum (bottom) of wild type BKPyV in the presence of 50-fold excess GD1b oligosaccharide. The spectrum is labeled as in C.

Mentions: BKPyV attachment is mediated by cell-surface sialic acid [8]. The most common sialic acid type in humans is 5-N-acetyl neuraminic acid (NeuNAc) [9]. Simians and most other mammals, however, possess an enzyme that can attach an additional hydroxyl group to NeuNAc, yielding 5-N-glycolyl neuraminic acid (NeuNGc). In contrast to humans, these animals therefore carry both NeuNAc and NeuNGc. Gangliosides were found to mediate cell attachment of BKPyV [10], and gangliosides GD1b and GT1b were later shown to function as specific receptors for BKPyV [11] (Fig. 1). Gangliosides are ceramide-based glycolipids, which are used as receptors for most of the well-characterized polyomaviruses, for example GD1a and GT1b for Polyoma, or GM1 for SV40 [12].


A structure-guided mutation in the major capsid protein retargets BK polyomavirus.

Neu U, Allen SA, Blaum BS, Liu Y, Frank M, Palma AS, Ströh LJ, Feizi T, Peters T, Atwood WJ, Stehle T - PLoS Pathog. (2013)

B-series gangliosides are receptors for BKPyV.(A) Schematic representation of structures of disialic acid-containing b-series gangliosides GD3, GD2, GD1b, GT1b and of the monosialylated a-series ganglioside GM1 indicating the abbreviated designations used here for individual residues in the gangliosides. (B) Ganglioside supplementation assays. Vero cells were incubated with gangliosides, challenged with BKPyV and scored for infection. The average number of VP1 positive cells is plotted compared to controls. Error bars represent the standard deviation for 3 independent experiments. Asterisks indicate p-value (*p<0.05). (C) STD off-resonance (top) and difference (bottom) spectra of WT BKPyV VP1 in the presence of 50-fold excess GD3 oligosaccharide. The off-resonance spectrum was scaled to 3%. GD3 resonances labeled in the difference spectrum receive considerable saturation transfer from the protein. Regions with strong signal overlap are not labeled because saturation effects in this region cannot be unambiguously assigned. Signals that were truncated are denoted by diagonal bars. (D) STD off-resonance (top) and difference spectrum (bottom) of wild type BKPyV in the presence of 50-fold excess GD1b oligosaccharide. The spectrum is labeled as in C.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3795024&req=5

ppat-1003688-g001: B-series gangliosides are receptors for BKPyV.(A) Schematic representation of structures of disialic acid-containing b-series gangliosides GD3, GD2, GD1b, GT1b and of the monosialylated a-series ganglioside GM1 indicating the abbreviated designations used here for individual residues in the gangliosides. (B) Ganglioside supplementation assays. Vero cells were incubated with gangliosides, challenged with BKPyV and scored for infection. The average number of VP1 positive cells is plotted compared to controls. Error bars represent the standard deviation for 3 independent experiments. Asterisks indicate p-value (*p<0.05). (C) STD off-resonance (top) and difference (bottom) spectra of WT BKPyV VP1 in the presence of 50-fold excess GD3 oligosaccharide. The off-resonance spectrum was scaled to 3%. GD3 resonances labeled in the difference spectrum receive considerable saturation transfer from the protein. Regions with strong signal overlap are not labeled because saturation effects in this region cannot be unambiguously assigned. Signals that were truncated are denoted by diagonal bars. (D) STD off-resonance (top) and difference spectrum (bottom) of wild type BKPyV in the presence of 50-fold excess GD1b oligosaccharide. The spectrum is labeled as in C.
Mentions: BKPyV attachment is mediated by cell-surface sialic acid [8]. The most common sialic acid type in humans is 5-N-acetyl neuraminic acid (NeuNAc) [9]. Simians and most other mammals, however, possess an enzyme that can attach an additional hydroxyl group to NeuNAc, yielding 5-N-glycolyl neuraminic acid (NeuNGc). In contrast to humans, these animals therefore carry both NeuNAc and NeuNGc. Gangliosides were found to mediate cell attachment of BKPyV [10], and gangliosides GD1b and GT1b were later shown to function as specific receptors for BKPyV [11] (Fig. 1). Gangliosides are ceramide-based glycolipids, which are used as receptors for most of the well-characterized polyomaviruses, for example GD1a and GT1b for Polyoma, or GM1 for SV40 [12].

Bottom Line: We have characterized the receptor specificity, structure and infectivity of the human polyomavirus BKPyV, the causative agent of polyomavirus-associated nephropathy, and uncover a molecular switch for binding different carbohydrate receptors.The crystal structure of the BKPyV capsid protein VP1 in complex with GD3 reveals contacts with two sialic acid moieties in the receptor, providing a basis for the observed specificity.Mutation of this residue from lysine in BKPyV to serine in SV40 switches the receptor specificity of BKPyV from GD3 to GM1 both in vitro and in cell culture.

View Article: PubMed Central - PubMed

Affiliation: Interfaculty Institute of Biochemistry, University of Tübingen, Tübingen, Germany.

ABSTRACT
Viruses within a family often vary in their cellular tropism and pathogenicity. In many cases, these variations are due to viruses switching their specificity from one cell surface receptor to another. The structural requirements that underlie such receptor switching are not well understood especially for carbohydrate-binding viruses, as methods capable of structure-specificity studies are only relatively recently being developed for carbohydrates. We have characterized the receptor specificity, structure and infectivity of the human polyomavirus BKPyV, the causative agent of polyomavirus-associated nephropathy, and uncover a molecular switch for binding different carbohydrate receptors. We show that the b-series gangliosides GD3, GD2, GD1b and GT1b all can serve as receptors for BKPyV. The crystal structure of the BKPyV capsid protein VP1 in complex with GD3 reveals contacts with two sialic acid moieties in the receptor, providing a basis for the observed specificity. Comparison with the structure of simian virus 40 (SV40) VP1 bound to ganglioside GM1 identifies the amino acid at position 68 as a determinant of specificity. Mutation of this residue from lysine in BKPyV to serine in SV40 switches the receptor specificity of BKPyV from GD3 to GM1 both in vitro and in cell culture. Our findings highlight the plasticity of viral receptor binding sites and form a template to retarget viruses to different receptors and cell types.

Show MeSH
Related in: MedlinePlus