Trans-complementation among naturally occurring deletion mutants of hepatitis B virus and integrated viral DNA for the production of viral particles with mutant genomes in hepatoma cell lines.
Cultured hepatoma cells (HepG2) were cotransfected with two different plasmids carrying a head-to-tail dimer of recombinant hepatitis B virus (HBV) DNA cloned from deletion mutants isolated from the circulation of persistently infected hosts. They were tested for the secretion of viral particles with mutant genome encapsidation. A recombinant plasmid defective in the S gene and one defective in both the C and P genes complemented in trans for the production of viral particles. Mutant genomes from either of the recombinants were encapsidated. Similarly, a recombinant defective in the C gene and another defective in the P gene trans-complemented for the production of viral particles containing mutant genomes. A hepatoma cell line with integrated HBV DNA sequences defective in the C and P genes (PLC/PRF/5) when transfected with a recombinant defective in the S gene produced viral particles with the HBV genome from the transfecting recombinants. These results confirm the expected trans-complementation among the S, C and P genes of HBV, when either episomal or integrated into chromosomes, for the maintenance of defective HBV mutants in persistently infected hosts.
Okamoto
H
H
Immunology Division, Jichi Medical School, Tochigi-Ken, Japan.
Wang
Y
Y
Tanaka
T
T
Machida
A
A
Miyakawa
Y
Y
Mayumi
M
M
D001483:Base Sequence; D006528:Carcinoma, Hepatocellular; D004279:DNA, Viral / Q000032:analysis; D017353:Gene Deletion; D005814:Genes, Viral / Q000235:genetics / Q000502:physiology*; D005816:Genetic Complementation Test; D016679:Genome, Viral; D006515:Hepatitis B virus / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation / Q000502:physiology*; D010957:Plasmids; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D014162:Transfection / Q000235:genetics; D014407:Tumor Cells, Cultured; D014771:Virion / Q000235:genetics
D004279:DNA, Viral; D011994:Recombinant Proteins