Anti-human immunodeficiency virus type 1 of a two-strand-system targeted to the polypurine tract

• Published in: International journal of molecular medicine Vol. 5; no. 2; p. 125
• Main Authors: Hiratou, T, Miyano-Kurosaki, N, Tsukahara, S, Tamura, Y, Takai, K, Yamamoto, N, Takaku, H
• Format: Journal Article
• Language: English
• Published: Greece 01.02.2000
• Subjects: Amino Acid Sequence; Anti-HIV Agents - chemistry; Anti-HIV Agents - pharmacology; Base Sequence; Cell Line; Chromatography, High Pressure Liquid; HIV-1 - chemistry; HIV-1 - drug effects; Humans; Microscopy, Fluorescence; Molecular Sequence Data; Oligonucleotides, Antisense - chemistry; Oligonucleotides, Antisense - pharmacology; Purines - chemistry; Reverse Transcriptase Inhibitors - chemistry; Reverse Transcriptase Inhibitors - pharmacology; Thionucleotides - chemistry; Thionucleotides - pharmacology
• ISSN: 1107-3756
• DOI: 10.3892/ijmm.5.2.125

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.