Characterization and structural localization of the reovirus lambda 3 protein.
The putative reovirus RNA polymerase, protein lambda 3, was characterized using antiserum prepared against a TrpE-lambda 3 fusion protein synthesized in Escherichia coli. Immunofluorescence microscopy showed that lambda 3 accumulated in perinuclear inclusion bodies in reovirus-infected cells. Analysis of lambda 3 accumulation in infected cells indicates that, once synthesized, lambda 3 is quite stable throughout the course of infection. Anti-lambda 3 serum did not immunoprecipitate virions, core particles or iodinated surface proteins of either virions or cores. These results indicate that lambda 3 is located in the inner part of the core. Experiments involving urea denaturation of purified reovirus cores indicate that lambda 3 cannot be selectively removed from the core without total denaturation of the core structure. When the dsRNA genome was eliminated from the core, lambda 3 remained associated with the other viral proteins in the core. Thus, lambda 3 appears to be a stable, structural component of the reovirus core, not bound to genomic dsRNA or free in soluble form inside the core.
Department of Microbiology, Medical College of Wisconsin, Milwaukee 53226.
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Cashdollar
D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology; D000918:Antibody Specificity; D000956:Antigens, Viral / Q000276:immunology; D002478:Cells, Cultured; D012321:DNA-Directed RNA Polymerases / Q000096:biosynthesis* / Q000276:immunology / Q000302:isolation & purification; D005347:Fibroblasts / Q000821:virology; D005455:Fluorescent Antibody Technique; D015151:Immunoblotting; D007553:Isotope Labeling; D012089:Mammalian orthoreovirus 3 / Q000187:drug effects / Q000201:enzymology / Q000254:growth & development*; D051379:Mice; D011233:Precipitin Tests; D012324:RNA-Dependent RNA Polymerase / Q000096:biosynthesis* / Q000276:immunology / Q000302:isolation & purification; D013462:Sulfur Radioisotopes; D013997:Time Factors; D014508:Urea / Q000494:pharmacology; D014764:Viral Proteins; D014771:Virion / Q000276:immunology; D014779:Virus Replication
D000914:Antibodies, Viral; D000956:Antigens, Viral; D013462:Sulfur Radioisotopes; D014764:Viral Proteins; D014508:Urea; C091731:lambda3 protein, Reovirus; D012324:RNA-Dependent RNA Polymerase; D012321:DNA-Directed RNA Polymerases