Structures of the Procapsid and Mature Virion of Enterovirus 71 Strain 1095

Cifuente, Javier O.; Lee, Hyunwook; Yoder, Jay A.; Shingler, Kristin L.; Carnegie, Michael S.; Yoder, Jennifer; Ashley, Robert E.; Makhov, Alexander M.; Conway, James F.; Hafenstein, Susan

doi:10.1128/jvi.03519-12

Cited by 37 publications

(37 citation statements)

References 63 publications

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“…3B), a region implicated in receptor binding and a virulence determinant (23). The basic, positively charged lysine at positon 244 was replaced with an acidic, negatively charged glutamic acid.…”

Section: Resultsmentioning

confidence: 99%

A Single Mutation in the VP1 of Enterovirus 71 Is Responsible for Increased Virulence and Neurotropism in Adult Interferon-Deficient Mice

Caine

Moncla

Ronderos

et al. 2016

J Virol

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Hand, foot, and mouth disease (HFMD) has spread throughout the Asia-Pacific region, affecting millions of young children, who develop symptoms ranging from painful blisters around their mouths and hands to neurological complications. Many members of the genus Enterovirus (family Picornaviridae) cause HFMD. Enterovirus 71 (EV71) is one of the primary causative agents and has been linked to severe disease. Vaccine efficacy and pathogenesis studies for EV71 have been limited because there is a lack of suitable animal models. Previously, we generated a mouse-adapted EV71 (mEV71) capable of infecting 12-week-old interferon receptor-deficient AG129 mice and used the model to evaluate the efficacy of candidate HFMD vaccines. Here, we present data investigating the genetic correlates of EV71 adaptation and characterize the virus's tissue tropism in mice. Using reverse genetics, a VP1 mutation (K244E) was shown to be necessary for mEV71 virulence in adult mice. Another VP1 mutation (H37R) was required for mEV71 recovery on rhabdomyosarcoma (RD) cells. Viral loads determined by real-time reverse transcription (RT)-PCR confirmed the presence of mEV71 in the sera and multiple organs of mice. Histological analysis revealed signs of meningitis and encephalitis, characteristic of severe human disease. The further description of this model has provided insight into EV71 pathogenesis and demonstrates the importance of the VP1 region in facilitating mEV71 adaptation. IMPORTANCEEV71 is a reemerging pathogen, and little is known about the genetic determinants involved in its pathogenesis. The absence of animal models has contributed to this lack of knowledge. The data presented here improve upon the existing animal models by characterizing a mouse-adapted strain of EV71. We determined that a VP1 mutation (K244E) was needed for EV71 virulence in adult AG129 mice. While this mutation was found previously for EV71 adaptation in 5-day-old BALB/c mice, neurotropic disease did not develop. Using interferon-deficient mice, we raised the age of susceptibility beyond 6 weeks and provided clear evidence that our model mimics severe human infections. The model can be exploited to identify determinants of EV71 virulence and to reveal molecular mechanisms that control the virus-host interaction, especially those associated with neurotropic disease. Furthermore, these data provide useful information regarding the importance of VP1, specifically position 244, in host adaptation and tissue dissemination. O ver the past decade, enterovirus 71 (EV71) (family Picornaviridae) was identified as one of the main causative agents responsible for large outbreaks of hand, foot, and mouth disease (HFMD) across the Asia-Pacific region. Other members of the genus Enterovirus cause HFMD, including coxsackieviruses A16, A6, A5, A7, A9, A10, B2, and B5, but EV71 has been linked to severe complications, including brainstem encephalitis, aseptic meningitis, and pulmonary edema (1-3). Picornaviruses, including the enteroviruses, have positive-sense, single-s...

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Section: Resultsmentioning

confidence: 99%

A Single Mutation in the VP1 of Enterovirus 71 Is Responsible for Increased Virulence and Neurotropism in Adult Interferon-Deficient Mice

Caine

Moncla

Ronderos

et al. 2016

J Virol

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“…Five residues that were mapped to capsid proteins were divergent between the 1095 strain (PDB accession no. 4GMP) (22) and our clinical C4 strain (Fig. 4), three of which are linked to EV71 antigenicity (34).…”

Section: Figmentioning

confidence: 99%

“…The structures of different strains of EV71 have been determined (20)(21)(22). By comparing the structures of the mature virion and a naturally occurring empty particle (sedimentation coefficient of 80S), Wang et al proposed a sensor-adaptor mechanism for EV71 uncoating (21) and found that the largest movement of the polypeptide occurs in residues 230 to 233 of VP1 at the end of the GH loop and at the beginning of strand H. They referred to this polypeptide region as the adaptor-sensor, which is directly downstream of a region of the GH loop that is external and structurally variable in poliovirus (21).…”

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confidence: 99%

Human Enterovirus 71 Uncoating Captured at Atomic Resolution

Lyu

Ding

Han

et al. 2014

J Virol

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Human enterovirus 71 (EV71) is the major causative agent of severe hand-foot-and-mouth diseases (HFMD) in young children, and structural characterization of EV71 during its life cycle can aid in the development of therapeutics against HFMD. Here, we present the atomic structures of the full virion and an uncoating intermediate of a clinical EV71 C4 strain to illustrate the structural changes in the full virion that lead to the formation of the uncoating intermediate prepared for RNA release. Although the VP1 N-terminal regions observed to penetrate through the junction channel at the quasi-3-fold axis in the uncoating intermediate of coxsackievirus A16 were not observed in the EV71 uncoating intermediate, drastic conformational changes occur in this region, as has been observed in all capsid proteins. Additionally, the RNA genome interacts with the N-terminal extensions of VP1 and residues 32 to 36 of VP3, both of which are situated at the bottom of the junction. These observations highlight the importance of the junction for genome release. Furthermore, EV71 uncoating is associated with apparent rearrangements and expansion around the 2-and 5-fold axes without obvious changes around the 3-fold axes. Therefore, these structures enabled the identification of hot spots for capsid rearrangements, which led to the hypothesis that the protomer interface near the junction and the 2-fold axis permits the opening of large channels for the exit of polypeptides and viral RNA, which is an uncoating mechanism that is likely conserved in enteroviruses. IMPORTANCE Human enterovirus 71 (EV71) is the major causative agent of severe hand-foot-and-mouth diseases (HFMD Human enterovirus 71 (EV71) is the leading causative agent for severe hand-foot-and-mouth diseases (HFMD) in infants and young children (1), and EV71 infection has caused significant morbidity and mortality in the Asia-Pacific regions. During 2008 to 2012, numerous outbreaks of HFMD occurred in mainland China, with over 2,000 fatal cases reported (www.chinacdc.cn). However, vaccines and effective drugs remain unavailable. EV71 belongs to Enterovirus species A within the Enterovirus genus of the Picornaviridae (2). The EV71 virion contains a singlestranded positive-sense RNA genome of 7.5 kb. The viral capsid, which has a diameter of approximately 300 Å, is composed of 60 copies each of VP1 to VP4 (3, 4) that are organized onto a quasi-Tϭ3 icosahedral lattice. The capsid proteins VP1, VP2, and VP3 each possess a ␤-sandwich jelly roll fold and form the outer surface of the capsid shell, whereas VP4 is situated inside the shell. The capsid surface is characterized by a depression encircling each 5-fold symmetry axis, which is referred to as the "canyon" and often contains the receptor-binding site in picornaviruses (5, 6). The hydrophobic pocket of VP1, which is located at the bottom of the canyon, contains a lipid moiety termed the "pocket factor," which likely stabilizes the mature virion. As observed in poliovirus and certain picornaviruses, receptor bindin...

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“…Recently, we and others have reported the structures of the virion, the naturally occurring empty particle, an uncoating intermediate or the A-particle, and the empty particle after the uncoating of EV71 as determined by x-ray crystallography or cryo-EM techniques (17)(18)(19)(20)(21). Despite its elusive role during the viral life cycle, the empty particle shares similar structural features with the virion and the uncoating intermediate.…”

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confidence: 99%

Crystal Structures of Enterovirus 71 (EV71) Recombinant Virus Particles Provide Insights into Vaccine Design

Lyu

Wang

et al. 2015

Journal of Biological Chemistry

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Background:No HFMD vaccine is available. Results: Structures of EV71 recombinant virus particles were determined showing that the inserted foreign peptide is exposed on the particle surface without capsid structural changes and that virus uncoating is not affected. Conclusion: VP1 BC loop is suitable for foreign peptide insertion for the generation of recombinant EV71 viruses. Significance: The results provide insights into vaccine development.

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Structures of the Procapsid and Mature Virion of Enterovirus 71 Strain 1095

Cited by 37 publications

References 63 publications

A Single Mutation in the VP1 of Enterovirus 71 Is Responsible for Increased Virulence and Neurotropism in Adult Interferon-Deficient Mice

A Single Mutation in the VP1 of Enterovirus 71 Is Responsible for Increased Virulence and Neurotropism in Adult Interferon-Deficient Mice

Human Enterovirus 71 Uncoating Captured at Atomic Resolution

Crystal Structures of Enterovirus 71 (EV71) Recombinant Virus Particles Provide Insights into Vaccine Design

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