Biosynthesis of reovirus-specified polypeptides: expression of reovirus S1-encoded sigma 1NS protein in transfected and infected cells as measured with serotype specific polyclonal antibody.
Polyclonal monospecific antibody was prepared against the reovirus serotype 1 Lang strain nonstructural sigma 1NS protein encoded by the S1 gene. The antibody was serotype-specific. The sigma 1NS protein of reovirus serotype 1, but not reovirus serotype 3, was recognized by the polyclonal antibody in both immunoprecipitation and Western immunoblot assays. The sigma 1NS protein expressed in vector-transfected COS cells was indistinguishable by immunoprecipitation and immunoblot analyses from the authentic sigma 1NS protein synthesized in virus-infected mouse L or monkey COS cells. The temporal appearance of sigma 1NS protein in virus-infected cells was similar to that of the other reovirus proteins. Both sigma 1NS and sigma 1, the two S1 gene products, were observed in the cytoplasm of COS cells by immunofluorescent microscopy, although their staining patterns were distinct from each other. However, sigma 1NS, but not sigma 1 or the other reovirion structural proteins, was also detected in the nucleoli of COS cells. These results suggest that sigma 1NS, like sigma 1, is a serotype-specific reovirus protein, but unlike sigma 1 is localized in part to the cell nucleus.
Belli
B A
BA
Department of Biological Sciences, University of California, Santa Barbara 93106.
Samuel
C E
CE
D000818:Animals; D000878:Anthranilate Synthase / Q000235:genetics / Q000378:metabolism; D015153:Blotting, Western; D002460:Cell Line; D002467:Cell Nucleus / Q000276:immunology; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay; D015870:Gene Expression / Q000502:physiology*; D005822:Genetic Vectors / Q000235:genetics; D000882:Haplorhini; D007700:Kinetics; D051379:Mice; D008856:Microscopy, Fluorescence; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D012087:Reoviridae / Q000378:metabolism*; D014162:Transfection / Q000235:genetics; D017361:Viral Nonstructural Proteins; D014764:Viral Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
D011993:Recombinant Fusion Proteins; D017361:Viral Nonstructural Proteins; D014764:Viral Proteins; C071901:sigma 1NS protein, Reovirus; D001616:beta-Galactosidase; D000878:Anthranilate Synthase