Expression of hepatitis B virus middle and large surface antigen genes in Saccharomyces cerevisiae.
The hepatitis B virus genome carries the surface antigen (SAg) gene and an open reading frame that encodes two SAg-related polypeptides: SAg with a 55-amino-acid N-terminal extension polypeptide and SAg with a 174-amino-acid N-terminal extension polypeptide. These are termed middle S and large S, respectively. These polypeptides or their glycosylated derivatives have been detected in Dane particles, but their chemical and biological properties have remained largely unknown because of their limited availability. We attempted to produce these proteins in Saccharomyces cerevisiae by placing the coding regions under the control of the promoter of the yeast glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene. Yeast cells carrying middle S and large S coding sequences produced 33,000- and 42,000-dalton products, respectively, each of which reacted with anti-S antibody and bound to polymerized human serum albumin, in accordance with the known properties of pre-S proteins from particles in human sera (K. H. Heermann, U. Goldmann, W. Schwartz, T. Seyffarth, H. Baumgarten, and W. H. Gerlich, J. Virol. 52:396-402, 1984; A. Machida, S. Kishimoto, H. Ohnuma, K. Baba, Y. Ito, H. Miyamoto, G. Funatsu, K. Oda, S. Usuda, S. Togami, T. Nakamura, Y. Miyakawa, and M. Mayumi, Gastroenterology 86:910-918, 1984). The middle S polypeptide is glycosylated and can be assembled into particles whose size and density are similar to those of SAg. However, this polypeptide was highly susceptible to proteolytic degradation into 29,000- and 26,000-dalton polypeptides, of which only the former retained the binding activity to polymerized albumin. The large S polypeptides are nonglycosylated, relatively stable, and do not seem to assemble into particles by themselves.
Imamura
T
T
Chemo-Sero-Therapeutic Research Institute, Kumamoto, Japan.
Araki
M
M
Miyanohara
A
A
Nakao
J
J
Yonemura
H
H
Ohtomo
N
N
Matsubara
K
K
D003001:Cloning, Molecular; D005796:Genes; D005814:Genes, Viral; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics; D006514:Hepatitis B Surface Antigens / Q000032:analysis / Q000235:genetics*; D006515:Hepatitis B virus / Q000235:genetics*; D010957:Plasmids; D011401:Promoter Regions, Genetic; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics*
D006514:Hepatitis B Surface Antigens; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases