Electron microscopy of hepatitis B virus core antigen expressing yeast cells by freeze-substitution fixation.
We have used the freeze-substitution fixation technique for electron microscopy of yeast cells that express the hepatitis B virus core antigen (HBcAg) following transformation with the cloned gene. Abundant spherical particles were found within the transformed cells. These particles had a uniform size and shape, measured about 21 nm in diameter, had electron-lucent centers, and consisted of many subunits. They were localized in both the cytoplasm and the nucleus. None of these particles was found in the cells of the parent strain. Comparison of the HBcAg particles isolated from the yeast cells and the particles within the yeast cells demonstrated that the 21-nm particles were in fact ultrastructurally superimposable on HBcAg. Thus, the HBcAg particles within the yeast cells were similar to the HBcAg particles in human liver tissues infected with hepatitis B virus, not only in their size and appearance, but also in their intracellular localization. These results suggest that the yeast cell has the same machinery for synthesis and intracellular translocation of the HBcAg polypeptides as the human cell.
Yamaguchi
M
M
Jikei University School of Medicine, Tokyo/Japan.
Hirano
T
T
Sugahara
K
K
Mizokami
H
H
Araki
M
M
Matsubara
K
K
D000818:Animals; D005615:Freezing; D005786:Gene Expression Regulation; D006512:Hepatitis B Core Antigens / Q000032:analysis / Q000235:genetics*; D006801:Humans; D008099:Liver / Q000473:pathology / Q000648:ultrastructure; D008854:Microscopy, Electron / Q000379:methods*; D011309:Preservation, Biological / Q000379:methods; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000648:ultrastructure; D014170:Transformation, Genetic
D006512:Hepatitis B Core Antigens