The Enterovirus 71 A-particle Forms a Gateway to Allow Genome Release: A CryoEM Study of Picornavirus Uncoating

Abstract

Since its discovery in 1969, enterovirus 71 (EV71) has emerged as a serious worldwide health threat. This human pathogen of the picornavirus family causes hand, foot, and mouth disease, and also has the capacity to invade the central nervous system to cause severe disease and death. Upon binding to a host receptor on the cell surface, the virus begins a two-step uncoating process, first forming an expanded, altered “A-particle”, which is primed for genome release. In a second step after endocytosis, an unknown trigger leads to RNA expulsion, generating an intact, empty capsid. Cryo-electron microscopy reconstructions of these two capsid states provide insight into the mechanics of genome release. The EV71 A-particle capsid interacts with the genome near the icosahedral two-fold axis of symmetry, which opens to the external environment via a channel ∼10 Å in diameter that is lined with patches of negatively charged residues. After the EV71 genome has been released, the two-fold channel shrinks, though the overall capsid dimensions are conserved. These structural characteristics identify the two-fold channel as the site where a gateway forms and regulates the process of genome release.

Author Summary

In a picornavirus capsid structural integrity must not be compromised until a key mechanism triggers genome release into a permissive cell. It has long been established that the majority of members of the picornavirus family solve this dilemma with a two-step uncoating process initiated by receptor recognition. For human enteroviruses, binding of an entry receptor triggers a series of conformational changes, resulting in an “A-particle” that is primed for genome release. After endocytosis, an unknown trigger causes the A-particle to expel the viral genome, leaving behind an emptied capsid. This process can be mimicked in solution by heating mature virus. Though the capsid species for both of these steps have been isolated, the fine details of the uncoating process have yet to be elucidated. Cryo-electron microscopy reconstructions of the enterovirus 71 A-particle and empty capsid provide compelling structural evidence to suggest that the icosahedral two-fold axis opens a channel that acts as a gateway in the viral capsid, regulating the release of genomic material from the altered particle.