Cryo-electron microscopy reconstructions of two types of wild rabbit hemorrhagic disease viruses characterized the structural features of Lagovirus

Hu, Zhide; Tian, Xiaojuan; Zhai, Yujia; Xu, Wei; Dong, Zheng; Sun, Fei

doi:10.1007/s13238-010-0007-0

Cited by 29 publications

(36 citation statements)

References 38 publications

(51 reference statements)

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“…Unfortunately, we were unable to obtain any crystals of RHDV suitable for X-ray diffraction owing to its propensity to degrade with time. From cryoEM micrographs ( Figure 1B ), consistent with previous observations [7], [22], two distinct classes of particles were observed: intact virions containing whole genomic RNA (high density inside) and “empty” virions containing sub-genomic RNA (low density inside). The presence of these two types of particles was confirmed by image classification ( Figure S1A ).…”

Section: Resultssupporting

confidence: 89%

“…Mature RHDV virions are spherical, non-enveloped particles with a T = 3, icosahedral capsid whose outer diameter varies between 32 and 44 nm and whose structure is defined by characteristic, cup-shaped depressions [6]. The only capsid protein present in RHDV, VP60, is composed of three domains, which include the N-terminal arm (NTA), the shell (S), and the protrusion (P), the latter of which is further divided into P1 and P2 sub-domains [7].…”

Section: Introductionmentioning

confidence: 99%

“…Previous structural studies of caliciviruses include three-dimensional (3D) cryo-electron microscopic (cryoEM) reconstructions of virus-like particles (VLPs) of Murine Norovirus (MNV, Norovirus ) and Feline calicivirus (FCV, Vesivirus ) at 8- and 16-Å resolution, respectively [10], [11], and determination of the crystal structures of the Norwalk virus (NV, Norovirus ) capsid at 3.4 Å [12], native FCV virions at 3.6 Å [13], and native virions of San Miguel sea lion virus (SMSV, Vesivirus ) at 3.2 Å [14]. CryoEM reconstructions of the RHDV VLP at 8 Å [15] and the native RHDV virion at 11 Å [7] have been computed and a C α homology model of RHDV was built based on the VLP cryo-reconstruction by using the crystal structure models of SMSV and FCV [16]. However, a more complete atomic model of RHDV is still lacking.…”

Section: Introductionmentioning

confidence: 99%

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Atomic Model of Rabbit Hemorrhagic Disease Virus by Cryo-Electron Microscopy and Crystallography

et al. 2013

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Rabbit hemorrhagic disease, first described in China in 1984, causes hemorrhagic necrosis of the liver. Its etiological agent, rabbit hemorrhagic disease virus (RHDV), belongs to the Lagovirus genus in the family Caliciviridae. The detailed molecular structure of any lagovirus capsid has yet to be determined. Here, we report a cryo-electron microscopic (cryoEM) reconstruction of wild-type RHDV at 6.5 Å resolution and the crystal structures of the shell (S) and protruding (P) domains of its major capsid protein, VP60, each at 2.0 Å resolution. From these data we built a complete atomic model of the RHDV capsid. VP60 has a conserved S domain and a specific P2 sub-domain that differs from those found in other caliciviruses. As seen in the shell portion of the RHDV cryoEM map, which was resolved to ∼5.5 Å, the N-terminal arm domain of VP60 folds back onto its cognate S domain. Sequence alignments of VP60 from six groups of RHDV isolates revealed seven regions of high variation that could be mapped onto the surface of the P2 sub-domain and suggested three putative pockets might be responsible for binding to histo-blood group antigens. A flexible loop in one of these regions was shown to interact with rabbit tissue cells and contains an important epitope for anti-RHDV antibody production. Our study provides a reliable, pseudo-atomic model of a Lagovirus and suggests a new candidate for an efficient vaccine that can be used to protect rabbits from RHDV infection.

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Section: Resultssupporting

confidence: 89%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Atomic Model of Rabbit Hemorrhagic Disease Virus by Cryo-Electron Microscopy and Crystallography

et al. 2013

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“…RHDV virions purified from infected rabbits consist of two particle types, intact and spikeless Tϭ3 capsids (26), highlighting the importance of S domain interactions. P domains in the Tϭ4 capsid nonetheless provide a distorted appearance, probably due to steric hindrance.…”

Section: Discussionmentioning

confidence: 99%

Epitope Insertion at the N-Terminal Molecular Switch of the Rabbit Hemorrhagic Disease Virus T=3 Capsid Protein Leads to Larger T=4 Capsids

Luque

González

Gómez-Blanco

et al. 2012

J Virol

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Viruses need only one or a few structural capsid proteins to build an infectious particle. This is possible through the extensive use of symmetry and the conformational polymorphism of the structural proteins. Using virus-like particles (VLP) from rabbit hemorrhagic disease virus (RHDV) as a model, we addressed the basis of calicivirus capsid assembly and their application in vaccine design. The RHDV capsid is based on a T=3 lattice containing 180 identical subunits (VP1). We determined the structure of RHDV VLP to 8.0-Å resolution by three-dimensional cryoelectron microscopy; in addition, we used San Miguel sea lion virus (SMSV) and feline calicivirus (FCV) capsid subunit structures to establish the backbone structure of VP1 by homology modeling and flexible docking analysis. Based on the three-domain VP1 model, several insertion mutants were designed to validate the VP1 pseudoatomic model, and foreign epitopes were placed at the N- or C-terminal end, as well as in an exposed loop on the capsid surface. We selected a set of T and B cell epitopes of various lengths derived from viral and eukaryotic origins. Structural analysis of these chimeric capsids further validates the VP1 model to design new chimeras. Whereas most insertions are well tolerated, VP1 with an FCV capsid protein-neutralizing epitope at the N terminus assembled into mixtures of T=3 and larger T=4 capsids. The calicivirus capsid protein, and perhaps that of many other viruses, thus can encode polymorphism modulators that are not anticipated from the plane sequence, with important implications for understanding virus assembly and evolution.

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“…Currently, structures of VLPs or virion particles of caliciviruses have been determined for GI.1 and GII.10 human NoVs (43,44), GV MNV (45,46), San Miguel sea lion virus (47), Tulane virus (48), rabbit hemorrhagic disease virus (49), and FCV (50). Therefore, structures are available for all classified genera of the family Caliciviridae, except for the genera Sapovirus and Nebovirus.…”

Section: Discussionmentioning

confidence: 99%

Mechanism of Cell Culture Adaptation of an Enteric Calicivirus, the Porcine Sapovirus Cowden Strain

Yokoyama

Chen

et al. 2016

J Virol

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The porcine sapovirus (SaV) (PoSaV) Cowden strain is one of only a few culturable enteric caliciviruses. Compared to the wildtype (WT) PoSaV Cowden strain, tissue culture-adapted (TC) PoSaV has two conserved amino acid substitutions in the RNAdependent RNA polymerase (RdRp) and six in the capsid protein (VP1). By using the reverse-genetics system, we identified that 4 amino acid substitutions in VP1 (residues 178, 289, 324, and 328), but not the substitutions in the RdRp region, were critical for the cell culture adaptation of the PoSaV Cowden strain. The other two substitutions in VP1 (residues 291 and 295) reduced virus replication in vitro. Three-dimensional (3D) structural analysis of VP1 showed that residue 178 was located near the dimer-dimer interface, which may affect VP1 assembly and oligomerization; residues 289, 291, 324, and 328 were located at protruding subdomain 2 (P2) of VP1, which may influence virus binding to cellular receptors; and residue 295 was located at the interface of two monomeric VP1 proteins, which may influence VP1 dimerization. Although reversion of the mutation at residue 291 or 295 from that of the TC strain to that of the WT reduced virus replication in vitro, it enhanced virus replication in vivo, and the revertants induced higher-level serum and mucosal antibody responses than those induced by the TC PoSaV Cowden strain. Our findings reveal the molecular basis for PoSaV adaptation to cell culture. These findings may provide new, critical information for the cell culture adaptation of other PoSaV strains and human SaVs or noroviruses. IMPORTANCEThe tissue culture-adapted porcine sapovirus Cowden strain is one of only a few culturable enteric caliciviruses. We discovered that 4 amino acid substitutions in VP1 (residues 178, 289, 324, and 328) were critical for its adaptation to LLC-PK cells. Two substitutions in VP1 (residues 291 and 295) reduced virus replication in vitro but enhanced virus replication and induced higherlevel serum and mucosal antibody responses in gnotobiotic pigs than those induced by the tissue culture-adapted strain. Structural modeling analysis of VP1 suggested that residue 178 may affect VP1 assembly and oligomerization; residues 289, 291, 324, and 328 may influence virus binding to cellular receptors; and residue 295 may influence VP1 dimerization. Our findings will provide new information for the cell culture adaptation of other sapoviruses and possibly noroviruses. C aliciviruses, in the family Caliciviridae, are small, icosahedral, and nonenveloped viruses with a diameter of 27 to 35 nm, which have a positive-sense, single-stranded RNA genome of 6.5 to 8.3 kb (1, 2). Caliciviruses have been classified into five genera (Norovirus, Sapovirus, Vesivirus, Lagovirus, and Nebovirus) and several proposed genera (3, 4). Among them, noroviruses (NoVs) and sapoviruses (SaVs) are the leading causes of gastroenteritis in humans of all ages. SaVs are often associated with sporadic, selflimiting gastroenteritis, the severity of which is reportedly milder t...

show abstract

Cryo-electron microscopy reconstructions of two types of wild rabbit hemorrhagic disease viruses characterized the structural features of Lagovirus

Cited by 29 publications

References 38 publications

Atomic Model of Rabbit Hemorrhagic Disease Virus by Cryo-Electron Microscopy and Crystallography

Atomic Model of Rabbit Hemorrhagic Disease Virus by Cryo-Electron Microscopy and Crystallography

Epitope Insertion at the N-Terminal Molecular Switch of the Rabbit Hemorrhagic Disease Virus T=3 Capsid Protein Leads to Larger T=4 Capsids

Mechanism of Cell Culture Adaptation of an Enteric Calicivirus, the Porcine Sapovirus Cowden Strain

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