Anti-HIV Activities and Mechanisms of Antisense Oligonucleotides

Polish - Japan Issue

Zeszyt specjalny. Polish - Japan Issue

We demonstrated that unmodified and modified (phosphorothioate) oligonucleotides preventcDNA synthesis by the AMV, MMLV, and HIV reverse transcriptases. Antisense oligonucleotide/RNA hybrids specifically arrest primer extension. The blockage involves the degradation ofthe RNA fragment bound to the antisense oligonucleotide by the reverse transcriptase associatedRNase H activity. However, the phosphorothioate oligomer inhibited polymerization by bindingto the AMV and MMLV RTs, rather than to the template RNA, whereas there was no competitivebinding of the phosphorothioate oligomer on the HIV RT during reverse transcription. Observation{of FlTC-S-ODN-reu-treated MOLT-4 cells with a confocal laser scanning microscope, revealedi diffuse fluorescence, apparently within the cytoplasm. Interestingly, fluorescent signals were; accumulated in the nuclear region of chronically infected MOLT-4/H1V-1 after a 60 min incubation. We also describe the long-term treatment of human immunodeficiency virus-infectedcells with antisense phosphorothioate oligonucleotides.