The ultrastructure of the developing replication site in foot-and-mouth disease virus-infected BHK-38 cells
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Paul Monaghan1, Hannah Cook1, Terry Jackson1
1Institute for Animal Health, Ash Road, Pirbright, Woking, Surrey GU24 0NF, UK.
Abstract:
Foot-and-mouth disease virus (FMDV) is the type species of the Aphthovirus genus of the Picornaviridae: Infection by picornaviruses results in a major rearrangement of the host cell membranes to create vesicular structures where virus genome replication takes place. In this report, using fluorescence and electron microscopy, membrane rearrangements in the cytoplasm of FMDV-infected BHK-38 cells are documented. At 1.5-2.0 h post-infection, free ribosomes, fragmented rough endoplasmic reticulum, Golgi and smooth membrane-bound vesicles accumulated on one side of the nucleus. Newly synthesized viral RNA was localized to this region of the cell. The changes seen in FMDV-infected cells distinguish this virus from other members of the Picornaviridae, such as poliovirus. Firstly, the collapse of cellular organelles to one side of the cell has not previously been observed for other picornaviruses. Secondly, the membrane vesicles, induced by FMDV, appear distinct from those induced by other picornaviruses such as poliovirus and echovirus 11 since they are relatively few in number and do not aggregate into densely packed clusters. Additionally, the proportion of vesicles with double membranes is considerably lower in FMDV-infected cells. These differences did not result from the use of BHK-38 cells in this study, as infection of these cells by another picornavirus, bovine enterovirus (a close relative of poliovirus), resulted in morphological changes similar to those reported for poliovirus-infected cells. With conventional fixation, FMDV particles were not seen; however, following high-pressure freezing and freeze-substitution, many clusters of virus-like particles were seen.
Insights
Foot-and-mouth disease virus (FMDV) causes unique cell membrane rearrangements, distinct from other picornaviruses. These changes involve organelle collapse and fewer, non-aggregated vesicles, aiding viral RNA replication.
Area of Science:
- Virology
- Cell Biology
- Molecular Biology
Background:
- Foot-and-mouth disease virus (FMDV) belongs to the Aphthovirus genus within the Picornaviridae family.
- Picornavirus infections induce host cell membrane rearrangements for viral genome replication.
Purpose of the Study:
- To document and characterize membrane rearrangements in FMDV-infected BHK-38 cells.
- To compare FMDV-induced changes with those caused by other picornaviruses.
Main Methods:
- Fluorescence microscopy
- Electron microscopy
- High-pressure freezing and freeze-substitution
Main Results:
- FMDV infection caused cellular organelles and ribosomes to accumulate on one side of the nucleus, with newly synthesized viral RNA localized to this region.
- FMDV-induced membrane vesicles were fewer in number, did not form dense clusters, and had a lower proportion of double membranes compared to poliovirus or echovirus 11.
- High-pressure freezing and freeze-substitution revealed clusters of virus-like particles, which were not visible with conventional fixation.
Conclusions:
- FMDV induces distinct cellular morphological changes compared to other picornaviruses like poliovirus.
- The observed membrane rearrangements are specific to FMDV and facilitate viral RNA replication.
- Advanced cryo-fixation techniques are crucial for visualizing picornavirus particles within infected cells.
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