Structural Study of Enterovirus Replication in Situ
- Project type: GACR EXPRO
- Project coordinator: CEITEC MU
- Grant provider/investor: Czech Science Foundation
- Project code: 19-25982X
- Implementation period: 01. 01. 2019 – 31. 12. 2023
- Budget: 1 872 761 EUR
Viruses from the genus Enterovirus, including coxsackieviruses and rhinoviruses, cause billions of infections every year. However, no anti-enterovirus treatments are available. We aim to structurally characterize enterovirus replication in situ to enable the identification of macromolecular interactions that could be targeted by therapeutics. We will use focused ion beam milling and cryo-electron tomography to determine: (1) how enteroviruses deliver their genomes into the cell cytoplasm; (2) whether enterovirus replication “factories” in infected cells contain the multi-subunit complexes of RNA-polymerases that were observed in vitro; and (3) whether enterovirus capsids assemble around genomes or if the genomes are packaged into capsids. To facilitate the cryo-electron tomography studies we will develop: (1) patterned grids to enable automation of the focused ion beam milling of cell sections; and (2) a method to embed macromolecules in rod-shaped vitreous ice to allow the recording of more complete tomographic data than is possible with currently used slab-shaped samples.