title: Effects of viscogens on RNA transcription inside reovirus particles.
abstract: The dsRNA genome of mammalian reovirus (MRV), like the dsDNA genomes of herpesviruses and many bacteriophages, is packed inside its icosahedral capsid in liquid-crystalline form, with concentrations near or more than 400 mg/ml. Viscosity in such environments must be high, but the relevance of viscosity for the macromolecular processes occurring there remains poorly characterized. Here, we describe the use of simple viscogens, glycerol and sucrose, to examine their effects on RNA transcription inside MRV core particles. Transcription inside MRV cores was strongly inhibited by these agents and to a greater extent than either predicted by theory or exhibited by a nonencapsidated transcriptase, suggesting that RNA transcription inside MRV cores is unusually sensitive to viscogen effects. The elongation phase of transcription was found to be a primary target of this inhibition. Similar results were obtained with particles of a second dsRNA virus, rhesus rotavirus, from a divergent taxonomic subfamily. Polymeric viscogens such as polyethylene glycol also inhibited RNA transcription inside MRV cores, but in a size-limited manner, suggesting that diffusion through channels in the MRV core is required for their activity. Modeling of the data suggested that the inherent intracapsid viscosity of both reo- and rotavirus is indeed high, two to three times the viscosity of water. The capacity for quantitative comparisons of intracapsid viscosity and effects of viscogens on macromolecular processes in confined spaces should be similarly informative in other systems.
authors: Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, Massachusetts 02115, USA.
authors: Aleksander A
authors: AA
authors: Demidenko
authors: Jinkee
authors: J
authors: Lee
authors: Thomas R
authors: TR
authors: Powers
authors: Max L
authors: ML
authors: Nibert
mesh_terms: D000818:Animals; D002460:Cell Line; D003451:Cryoprotective Agents / Q000494:pharmacology*; D016679:Genome, Viral / Q000502:physiology; D005990:Glycerol / Q000494:pharmacology*; D006801:Humans; D051379:Mice; D012330:RNA, Double-Stranded / Q000378:metabolism*; D012367:RNA, Viral / Q000096:biosynthesis*; D012087:Reoviridae / Q000378:metabolism*; D013395:Sucrose / Q000494:pharmacology*; D013549:Sweetening Agents / Q000494:pharmacology*; D014158:Transcription, Genetic / Q000187:drug effects*; D014783:Viscosity / Q000187:drug effects; D014867:Water / Q000378:metabolism
chemical_list: D003451:Cryoprotective Agents; D012330:RNA, Double-Stranded; D012367:RNA, Viral; D013549:Sweetening Agents; D014867:Water; D013395:Sucrose; D005990:Glycerol