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. The use of electron microscope tomography revealed the membrane structure and topology of nodavirus RNA replication compartments. A universal feature of positive-strand RNA viruses is the involvement of host intracellular membranes in RNA replication. Intracellular membrane rearrangements associated with viral RNA replication have been observed for many positive-strand RNA viruses including alphaviruses, picornaviruses, flaviviruses, and others. In the nodavirus flock house virus (FHV), viral RNA synthesis occurs on mitochondrial membranes in association with formation of 40 60 nm vesicles or spherules between the inner and outer mitochondrial membranes. For brome mosaic virus in the alphavirus superfamily, RNA replication has been found to occur inside strikingly similar spherules on endoplasmic reticulum membranes. To better define the structure and topology of such viral RNA replication structures, we studied the spherules formed in FHV-infected Drosophila cells by electron microscope tomography, which permits the visualization of cellular structures in three dimensions.
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