Targets of a protease inhibitor, KNI-272, in HIV-1-infected cells.
The targets of a protease inhibitor, KNI-272, in the HIV-1 life cycle were investigated in this study. Neither expression of HIV-1 Gag proteins nor production of virus particles was detected in cells infected acutely with HIV-1 cultured in the presence of KNI-272. Although HIV-1 proviral DNA was detected in the cells by PCR, the inhibitor depressed the amount of the proviral DNA in a concentration dependent manner. These results indicate that one of the targets of KNI-272 occurs in the stage before the expression of viral structural proteins. No direct inhibition of reverse transcription was found with the inhibitor. To confirm the inhibition of viral protease, persistently HIV-1-infected cells were cultured in the presence of the inhibitor and examined by electron microscopy for the morphology of HIV-1 particles. Doughnut-shaped immature particles were observed in the extracellular space of the cells, and disrupted semicircular shaped particles were also seen at the higher concentration of KNI-272. A bioassay for infectivity showed that the virus particles were not infectious, and immunofluorescent assay using anti-p17 antibody, that does not react with the precursor of Gag protein, revealed that Gag precursor p55 protein in the cells was not processed. Thus, KNI-272 blocked the maturation of viral particles. Consequently, KNI-272 has at least two inhibition targets in the stages of the HIV-1 life cycle.
Goto
T
T
Department of Microbiology, Osaka Medical College, Takatsuki, Osaka, Japan. tgoto@art.osaka-med.ac.jp
Nakano
T
T
Kohno
T
T
Morimatsu
S
S
Morita
C
C
Hong
W
W
Kiso
Y
Y
Nakai
M
M
Sano
K
K
D019380:Anti-HIV Agents / Q000494:pharmacology* / Q000633:toxicity; D015496:CD4-Positive T-Lymphocytes / Q000187:drug effects / Q000821:virology; D002460:Cell Line; D002478:Cells, Cultured; D017320:HIV Protease Inhibitors / Q000494:pharmacology* / Q000633:toxicity; D015497:HIV-1 / Q000187:drug effects* / Q000235:genetics / Q000254:growth & development / Q000648:ultrastructure; D009842:Oligopeptides / Q000494:pharmacology* / Q000633:toxicity; D014176:Protein Biosynthesis / Q000187:drug effects; D019065:Virus Assembly / Q000187:drug effects
D019380:Anti-HIV Agents; D017320:HIV Protease Inhibitors; D009842:Oligopeptides; C077822:kynostatin 272