MEMBRANE BOUND RNA VIRUS REPLICATION COMPLEXES

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This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. RNA replication by positive-strand RNA viruses is universally associated with membranes and membrane rearrangements such as single or double membrane vesicles. However, in most cases the site(s) of RNA synthesis relative to these rearranged membranes and the topological relationships between membranes, vesicle interiors, lumenal compartments and the cytoplasm are poorly defined. Recent results suggest that, in at least some cases, viral RNA replication occurs inside such vesicles. However, these findings leave many unresolved questions, in many ways due to limitations inherent in the two-dimensional analyses used. The best-studied member of the nodavirus family, flock house virus (FHV), replicates at the outer mitochondrial membranes of infected Drosophila cells where membrane-bound spherules form in the intermembrane space, similar to multi-vesicular structures observed for many positive-strand RNA viruses. We employ electron microscope tomography (EMT) to resolve the morphology of the FHV spherule membranes.