Replication-dependent 65R→K reversion in human immunodeficiency virus type 1 reverse transcriptase double mutant K65R + L74V
| Autor: | Christina Amat di San Filippo, Sara Skaggs, Raymond F. Schinazi, Viktoria Nurpeisov, Prem L. Sharma, Kimberly Lee |
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| Rok vydání: | 2004 |
| Předmět: |
Mutant
Reversion Proviral DNA Mutagenesis (molecular biology technique) Biology Arginine Virus Replication Virus Nucleoside Reverse Transcriptase Inhibitor 03 medical and health sciences Virology Viral replication Humans Point Mutation Cells Cultured 030304 developmental biology Human immunodeficiency virus type 1 (HIV-1) 0303 health sciences Guanosine 030306 microbiology Point mutation Lysine Dioxolanes Molecular biology Human interleukin-2 (IL-2) Reverse transcriptase HIV Reverse Transcriptase 3. Good health Drug Design Drug resistance HIV-1 Leukocytes Mononuclear Reverse Transcriptase Inhibitors Replication fitness |
| Zdroj: | Virology. 321(2):222-234 |
| ISSN: | 0042-6822 |
| DOI: | 10.1016/j.virol.2003.11.013 |
| Popis: | Understanding of the mechanisms of interaction among nucleoside reverse transcriptase inhibitor (NRTI)-selected mutations in the human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) coding sequence is essential for the design of newer drugs and for enhancing our vision of the structure function relationship among amino acids of the polymerase domain of HIV-1. Although several nucleoside reverse transcriptase inhibitors select RT mutations K65R and L74V, the combination of 65R + 74V is rare in clinics. A novel NRTI (−)-β-d-dioxolane-guanosine (DXG) is known to select in vitro either the 65R or 74V mutant virus (Antimicrob. Agents Chemother. 44 (2000) 1783). These mutations were not selected together during repeated passaging of the HIV-1 in the presence of this drug. To analyze the impact of these RT mutations on viral replication, a double mutant containing K65R + L74V was created by site-directed mutagenesis in a pNL4-3 background. Replication kinetic assays revealed that the mutant K65R + L74V is unstable, and 65R→K reversion occurs during replication of virus in phytohemagglutinin (PHA)-stimulated human peripheral blood mononuclear (PBM) cells in the absence of selection pressure. Replication kinetic assays in MT-2 cells demonstrated that double mutant 65R + 74V is highly attenuated for replication and the initiation of reversion is related to the increase in RT activity. Additionally, the suppression of viral replication in the presence of DXG or under suboptimal human recombinant interleukin-2 leads to minimal or no 65R→K reversion. These observations provide evidence that 65R→K reversion in the double mutant 65R + 74V is dependent on a specific rate of viral replication in a pNL4-3 background. A similar phenomenon may occur in vivo, which may have implications for treatment management strategies. |
| Databáze: | OpenAIRE |
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