Inhibition of HIV-1 reverse transcription by triple-helix forming oligonucleotides with viral RNA.
Reverse transcription of retroviral RNA into double-stranded DNA is catalyzed by reverse transcriptase (RT). A highly conserved polypurine tract (PPT) on the viral RNA serves as primer for plus-strand DNA synthesis and is a possible target for triple-helix formation. Triple-helix formation during reverse transcription involves either single-stranded RNA or an RNA.DNA hybrid. The effect of triple-helix formation on reverse transcription has been analyzed here in vitro using a three-strand-system consisting of an RNA.DNA hybrid and triplex-forming oligonucleotides (TFOs) consisting either of DNA or RNA. Three strand triple-helices inhibit RNase H cleavage of the PPT-RNA.DNA hybrid and initiation of plus-strand DNA synthesis in vitro. Triple-helix formation on a single-stranded RNA target has also been tested in a two-strand-system with TFOs comprising Watson-Crick and Hoogsteen base-pairing sequences, both targeted to the PPT-RNA, on a single strand connected by a linker (T)4. TFOs prevent RNase H cleavage of the PPT-RNA and initiation of plus-strand DNA synthesis in vitro. In cell culture experiments one TFO is an efficient inhibitor of retrovirus replication, leading to a block of p24 synthesis and inhibition of syncytia formation in newly infected cells.
Volkmann
S
S
Max-Planck-Institut für Molekulare Genetik, Abt. Schuster, Berlin Dahlem, Germany.
Jendis
J
J
Frauendorf
A
A
Moelling
K
K
D000998:Antiviral Agents / Q000737:chemistry / Q000494:pharmacology; D001483:Base Sequence; D002460:Cell Line; D002472:Cell Transformation, Viral; D054303:HIV Reverse Transcriptase; D015497:HIV-1 / Q000187:drug effects / Q000235:genetics* / Q000502:physiology; D006801:Humans; D066298:In Vitro Techniques; D008954:Models, Biological; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D009841:Oligonucleotides / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology*; D012367:RNA, Viral / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology; D012194:RNA-Directed DNA Polymerase / Q000235:genetics; D016914:Ribonuclease H; D014158:Transcription, Genetic; D014779:Virus Replication / Q000187:drug effects
D000998:Antiviral Agents; D009841:Oligonucleotides; D012367:RNA, Viral; D054303:HIV Reverse Transcriptase; D012194:RNA-Directed DNA Polymerase; D016914:Ribonuclease H