Expression of coronavirus E1 and rotavirus VP10 membrane proteins from synthetic RNA.
Some viruses acquire their envelopes by budding through internal membranes of their host cell. We have expressed the cloned cDNA for glycoproteins from two such viruses, the E1 protein of coronavirus, which buds in the Golgi region, and VP10 protein of rotavirus, which assembles in the endoplasmic reticulum. Messenger RNA was prepared from both cDNAs by using SP6 polymerase and either translated in vitro or injected into cultured CV1 cells or Xenopus oocytes. In CV1 cells, the E1 protein was localised to the Golgi region and VP10 protein to the endoplasmic reticulum. In Xenopus oocytes, the E1 protein acquired post-translational modifications indistinguishable from the sialylated, O-linked sugars found on viral protein, while the VP10 protein acquired endoglycosidase-H-sensitive N-linked sugars, consistent with their localisation to the Golgi complex and endoplasmic reticulum, respectively. Thus the two proteins provide models with which to study targeting to each of these intracellular compartments. When the RNAs were expressed in matured, meiotic oocytes, the VP10 protein was modified as before, but the E1 protein was processed to a much lesser extent than in interphase oocytes, consistent with a cessation of vesicular transport during cell division.
D000818:Animals; D002460:Cell Line; D003332:Coronaviridae / Q000235:genetics*; D004721:Endoplasmic Reticulum; D006056:Golgi Apparatus; D008954:Models, Biological; D009865:Oocytes / Q000378:metabolism; D010957:Plasmids; D014176:Protein Biosynthesis; D012313:RNA; D012401:Rotavirus / Q000235:genetics*; D014170:Transformation, Genetic; D014763:Viral Matrix Proteins / Q000096:biosynthesis / Q000235:genetics*; D014981:Xenopus
D014763:Viral Matrix Proteins; D012313:RNA