Investigation of the Nodavirus lifecycle in vivo

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DESCRIPTION (provided by applicant): Flock House Virus (FHV) is a non-enveloped, icosahedral RNA virus that infects insects. FHV has been thoroughly studied genetically, biophysically, and structurally, which makes it an excellent system for extending these studies to investigate the virus lifecycle (virus entry, disassembly, assembly, and cellular exit) in situ. To study the FHV lifecycle in situ the virus coat protein containing the tetracysteine tag will be labeled specifically with biarsenical fluorophores, which are visible in both the fluorescent and electron microscopes allowing the different images to be correlated. Additionally, in optical pulse-chase experiments the FHV will be successively labeled with different colored biarsenical fluorophores to distinguish older and younger coat protein. To investigate at higher resolution the spatial relationship of FHV within cells, tomographic electron microscope images will be used to produce three-dimensional reconstructions of the virus infected cells.