Encapsidation of truncated human hepatitis B virus genomes through trans-complementation of the core protein and polymerase.
Mutational analyses and complementation tests were used to analyze the strategy of packaging and of replication of human hepatitis B virus (HBV). By creating new restriction enzyme sites and by varying the genome length of HBV mutants, we identified that the mutated genomes could be encapsidated through trans-complementation of the polymerase and/or core protein. This study demonstrates that the polymerase of HBV, similar to that of duck hepatitis B virus (DHBV), is synthesized de novo instead of through a core-polymerase fusion protein. The results also indicate that both the polymerase and the core protein can be supplied in trans during viral packaging, and that the complementation is not due to recombination between the cotransfected plasmids. Furthermore, HBV genome deleted down to 2.4 kb is still able to be encapsidated, as measured by the endogenous polymerase reaction. Taken together, these results provide a basis for using HBV as a vector to deliver foreign genes into hepatocytes and for defining the location of the packaging signal on the HBV genome.
Chiang
P W
PW
Department of Medical Research, Veterans General Hospital, Taipei, Taiwan, Republic of China.
Hu
C P
CP
Su
T S
TS
Lo
S C
SC
Chu
M H
MH
Schaller
H
H
Chang
C M
CM
D002213:Capsid / Q000378:metabolism*; D002460:Cell Line; D003001:Cloning, Molecular; D004252:DNA Mutational Analysis; D004259:DNA-Directed DNA Polymerase / Q000235:genetics* / Q000378:metabolism; D005814:Genes, Viral; D005816:Genetic Complementation Test; D006515:Hepatitis B virus / Q000201:enzymology / Q000235:genetics* / Q000502:physiology; D006801:Humans; D010957:Plasmids; D015183:Restriction Mapping; D014162:Transfection; D014758:Viral Core Proteins / Q000235:genetics* / Q000378:metabolism; D014779:Virus Replication
D014758:Viral Core Proteins; D004259:DNA-Directed DNA Polymerase