Anti-human immunodeficiency virus type 1 of a two-strand-system targeted to the polypurine tract.
The polypurine tract (PPT) is highly conserved among the known human immunodeficiency virus (HIV)-1 strains, and is a possible target for triplex formation. We show the effects of triple-helix formation by assays of primer extension inhibition in vitro, using a two-strand-system (FTFOs) targeted to the PPT of HIV-1. The two-stranded composition of a triple-helix is thermodynamically and kinetically superior to the three-strand-system. The FTFOs inhibited the RT activity in a sequence-specific manner, i.e., the triplex actually formed at the PPT and blocked the RT. The FTFOs containing the phosphorothioate groups at the antisense sequences showed greater 3'-exonuclease resistance. In the observation of the FITC-DsDGloopT5-37 with MOLT-4 cells by a confocal laser scanning microscope, diffuse fluorescence was apparently observed in the cytoplasm and nucleus. However, weak fluorescence was observed within the cytoplasm and nucleus of MOLT-4 cells treated with the antisense phosphorothioate oligonucleotides (S-ODN-gag-AUG). In HIV-1 infected MOLT-4 cells, the FTFOs containing the phosphorothioate groups at the antisense sequence sites and guanosine rich parts within the third Hoogsteen base pairing sequence inhibit the replication of HIV-1 more effectively than the antisense oligonucleotides, indicating sequence-specific inhibition of HIV-1 replication.
Hiratou
T
T
Department of Industrial Chemistry, Chiba Institute of Technology, Narashino, Chiba 275-0016, Japan.
Miyano-Kurosaki
N
N
Tsukahara
S
S
Tamura
Y
Y
Takai
K
K
Yamamoto
N
N
Takaku
H
H
D000595:Amino Acid Sequence; D019380:Anti-HIV Agents / Q000737:chemistry* / Q000494:pharmacology; D001483:Base Sequence; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid; D015497:HIV-1 / Q000737:chemistry* / Q000187:drug effects; D006801:Humans; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D016376:Oligonucleotides, Antisense / Q000737:chemistry* / Q000494:pharmacology; D011687:Purines / Q000737:chemistry*; D018894:Reverse Transcriptase Inhibitors / Q000737:chemistry* / Q000494:pharmacology; D013873:Thionucleotides / Q000737:chemistry* / Q000494:pharmacology
D019380:Anti-HIV Agents; D016376:Oligonucleotides, Antisense; D011687:Purines; D018894:Reverse Transcriptase Inhibitors; D013873:Thionucleotides