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Comparative Study
. 2017 Jan;98(1):68-76.
doi: 10.1099/jgv.0.000658. Epub 2017 Feb 6.

Vesivirus 2117 capsids more closely resemble sapovirus and lagovirus particles than other known vesivirus structures

Michaela Conley  1 , Edward Emmott  2 , Richard Orton  1 , David Taylor  3   4 , Daniel G Carneiro  5   1 , Kazuyoshi Murata  4 , Ian G Goodfellow  2 , Grant S Hansman  6   4 , David Bhella  1
Affiliations
Comparative Study

Vesivirus 2117 capsids more closely resemble sapovirus and lagovirus particles than other known vesivirus structures

Michaela Conley et al. J Gen Virol. 2017 Jan.

Abstract

Vesivirus 2117 is an adventitious agent that, in 2009, was identified as a contaminant of Chinese hamster ovary cells propagated in bioreactors at a pharmaceutical manufacturing plant belonging to Genzyme. The consequent interruption in supply of Fabrazyme and Cerezyme (drugs used to treat Fabry and Gaucher diseases, respectively) caused significant economic losses. Vesivirus 2117 is a member of the Caliciviridae, a family of small icosahedral viruses encoding a positive-sense RNA genome. We have used cryo-electron microscopy and three-dimensional image reconstruction to calculate a structure of vesivirus 2117 virus-like particles as well as feline calicivirus and a chimeric sapovirus. We present a structural comparison of several members of the Caliciviridae, showing that the distal P domain of vesivirus 2117 is morphologically distinct from that seen in other known vesivirus structures. Furthermore, at intermediate resolutions, we found a high level of structural similarity between vesivirus 2117 and Caliciviridae from other genera: sapovirus and rabbit hemorrhagic disease virus. Phylogenetic analysis confirms vesivirus 2117 as a vesivirus closely related to canine vesiviruses. We postulate that morphological differences in virion structure seen between vesivirus clades may reflect differences in receptor usage.

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Figures

Fig. 1.
Fig. 1.
Cryo-EM structure of vesivirus 2117 at 10 Å resolution. (a) Cryo-electron micrograph of vesivirus 2117 VLPs imaged in a frozen-hydrated state. Bar, 100 nm. (b) A central slice through the 3D reconstruction of vesivirus 2117 shows the compact structure of the P domain. (c) Stereo pair images of the reconstruction, calculated at 10 Å resolution, viewed along the twofold symmetry axis. (d) A side view of the 2117 VP1 dimer viewed parallel to the capsid surface highlights the pronounced horn-shaped structures on the outer faces of the P domains.
Fig. 2.
Fig. 2.
3D reconstruction of a chimeric sapovirus VLP. (a) Cryo-electron micrograph of chimeric sapovirus-like particles. Bar, 100 nm. (b) A central slice through the sapovirus VLP structure reveals a compact P domain structure, similar to that seen in 2117. (c) Stereo pair images of the reconstruction calculated at 10 Å resolution, viewed along the twofold symmetry axis. (d) Side view of the sapovirus VP1 dimer.
Fig. 3.
Fig. 3.
Structure of FCV. (a) Cryo-microscopy of native FCV virions. Bar, 100 nm. (b) The central section shows a broader P domain structure that presents a flatter outer face. (c) Stereo pair images of the reconstruction, calculated at 7 Å resolution, viewed along the twofold symmetry axis. (d) A side view of a dimeric FCV VP1 capsomere.
Fig. 4.
Fig. 4.
Side-by-side comparison of (a) vesivirus 2117, (b) a chimeric sapovirus, (c) RHDV, (d) FCV, (e) norovirus GII.10 and (f) NoV at 10 Å resolution, viewed along the twofold symmetry axis.
Fig. 5.
Fig. 5.
Side-by-side comparison of the P domains of (a) vesivirus 2117, (b) a chimeric sapovirus, (c) RHDV, (d) FCV, (e) norovirus GII.10 and (f) NoV at 10 Å resolution.
Fig. 6.
Fig. 6.
Vesivirus 2117 neighbour-joining tree. The evolutionary history of the VP1 capsid of 2117 was inferred using the neighbour-joining method. The vesivirus 2117 cluster is highlighted in red; FCV cluster, in orange; VESV cluster (includes walrus, reptile and sea lion viruses), in green; and the sapovirus cluster, in blue. The proportions of replicate trees in which the associated taxa clustered together in the bootstrap test (1000 replicates) are shown next to the branches. The tree is drawn to scale, with branch lengths in the same units as those of the evolutionary distances used to infer the phylogenetic tree. The genus Vesivirus is highlighted in yellow; the genus Sapovirus, in blue; the genus Nebovirus, in dark grey; the genus Lagovirus, in light grey; and the genus Norovirus, in pink. BECV, bovine enteric calicivirus; BoNV, bovine norovirus; CCV, canine calicivirus; CNV, canine norovirus; CVV, canine vesivirus; EBHSV, European brown hare syndrome virus; FCV, feline calicivirus; HuCV, human calicivirus; HuNV-GI, human norovirus GI; HuNV-GII, human norovirus GII; MCV, mink calicivirus; MuNV, murine norovirus; PESV, porcine enteric sapovirus; RaCV, rabbit calicivirus; RaVV, rabbit vesivirus; ReCV, reptile calicivirus; ReVV, reptile vesivirus; RHDV, rabbit hemorrhagic disease virus; SMSV, San Miguel sea lion virus; SMSV1, San Miguel sea lion virus 1; SSLV, Steller sea lion vesivirus; SwNV, swine norovirus; VESV, vesicular exanthema of swine virus; WCV, walrus calicivirus.
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