Inhibition of HIV-1 ReplicationIn Vitroand in Human Infected Cells by Modified Antisense Oligonucleotides Targeting the tRNALys3/RNA Initiation Complex
| Autor: | Simon Litvak, Frédéric Freund, Justine Michel, Florence Boulmé, Michel Ventura, Serge Moreau |
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| Rok vydání: | 2001 |
| Předmět: |
Pharmacology
Base Sequence Transcription Genetic Functional features Ribonuclease H Human immunodeficiency virus (HIV) RNA Oligonucleotides Antisense Biology Virus Replication medicine.disease_cause Molecular biology Reverse transcriptase In vitro Antisense oligonucleotides HIV-1 Mutagenesis Site-Directed Genetics medicine Humans Nucleic Acid Conformation RNA Transfer Lys RNA Viral Primer binding site |
| Zdroj: | Antisense and Nucleic Acid Drug Development. 11:301-315 |
| ISSN: | 1087-2906 |
| Popis: | The untranslated 5' leader region of the human immunodeficiency virus type 1 (HIV-1) RNA plays an essential role in retroviral replication. It is the first retrotranscribed RNA region, primed from a cellular tRNALys3 partially annealed to the HIV-1 primer binding site (PBS). The structural and functional features of the HIV-1 reverse transcription initiation complex have been thoroughly studied. In this work, we used chemically modified antisense oligonucleotides (AS-ODN) as competitors of the natural tRNALys3 primer for the PBS region. Modified 2'-O-methyl AS-ODN were able to inhibit in vitro HIV-1 reverse transcription and displace the tRNALys3 previously annealed to the PBS. The destabilization of the initiation complex by 2'-O-methyl ODN was a sequence-specific process. We further demonstrated the importance of an anchor region contiguous to the PBS in the annealing of the antisense molecule, allowing the displacement of tRNALys3. The 20-mer 2'-O-methyl molecules were also able to inhibit viral replication in HIV-1-human infected cells, either by blocking cDNA synthesis during the early phase or by interfering with the annealing of the tRNALys3 primer to the PBS during the late phase of the viral cycle. Thus, the highly conserved retroviral initiation complex was shown to be a promising target when using the antisense strategy. |
| Databáze: | OpenAIRE |
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