Molecular mechanisms of enterovirus replication

Human enteroviruses cause a multitude of diseases ranging from common cold to acute paralysis and potentially life-threatening infections of the heart.

All enteroviruses share a common mode of replication inside the infected cell: cytoplasmic membranes are rapidly hijacked and reshaped into viral RNA factories, so-called replication complexes (RCs). The detailed organisation of RCs inside infected cells, as well as their link to virus particle assembly, is no known.

In this proposal we aim to combine novel experimental approaches to reach an integrated structural and mechanistic understanding of enterovirus replication.

Specifically, we will: (i) conduct in-cell structural studies using focussed ion beam-milling and cryo-electron tomography.

Preliminary data indicate that this approach, combined with pharmacological treatment and KO cells, can reveal several striking and distinct roles for different components of the autophagy pathway, at a resolution that allow identification of novel macromolecular complexes, (ii) determine the structure and macromolecular composition of a novel protein filament we have identified (preliminary data) in poliovirus-infected cells, (iii) reconstitute the membrane-bound, autophagy protein-assisted assembly of the poliovirus capsid.

We believe that the proposed experiments will vastly expand the structural and mechanistic understanding of enterovirus replication, which may aid in design of new antiviral strategies.