Nov 1, 1986

Expression of the RNA genome of an animal virus in Saccharomyces cerevisiae

Proceedings of the National Academy of Sciences of the United States of America
M MakarowL Kääriäinen


The nucleocapsid of vesicular stomatitis virus (VSV) was introduced into the cytoplasm of Saccharomyces cerevisiae by low pH-dependent fusion of the viral envelope with the spheroplast plasma membrane. This led to de novo synthesis of the three major structural proteins of the virus--the G, N, and M proteins--as shown by immunoprecipitation of [35S]methionine-labeled spheroplast lysates. In NaDodSO4/polyacrylamide gel electrophoresis, M and N proteins comigrated with those of the virion, whereas the yeast-made G protein migrated as two bands with apparent molecular sizes of 60 and 70 kDa. Both polypeptides appeared to be N-glycosylated, since only one polypeptide with the apparent molecular mass of approximately equal to 55 kDa was produced in the presence of tunicamycin. Phase separation into Triton X-114 suggested that the unglycosylated G protein was membrane bound. According to immunofluorescent surface staining of live spheroplasts, at least part of the G protein was transported to the plasma membrane. Spheroplasts expressing the VSV genes could be fused together by low pH to form polykaryons, indicating that G protein synthetized by yeast was fusogenic--i.e., biologically active.

  • References21
  • Citations10


  • References21
  • Citations10


Mentioned in this Paper

Corona Virus Membrane Protein E1
Viral Proteins
Genes, Viral
Vesicular stomatitis Indiana virus
Granulin Matrix Proteins
Membrane Glycoproteins
Vesicular Stomatitis

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