Mechanism of resistance of human immunodeficiency virus type 1 to 2',3'-dideoxyinosine.
- 1 July 1993
- journal article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences
- Vol. 90 (13), 6135-6139
- https://doi.org/10.1073/pnas.90.13.6135
Abstract
A molecular clone containing the wild-type reverse transcriptase (RT) coding region of human immunodeficiency virus type 1 (HIV-1) was constructed, and site-directed mutagenesis was used to introduce mutations--Leu74-->Val (L74V), T215Y, and the combination L74V/T215Y--into the RT coding region. The proteins were purified by immunoaffinity chromatography. Assays were performed with mutant and wild-type RT to determine substrate and inhibitor specificity. All three mutant enzymes catalyzed the incorporation of substrate 2'-deoxynucleoside 5'-triphosphates (dNTPs) as efficiently as wild-type HIV-1 RT. Small changes were observed in the Km values for dNTPs with all three mutant enzymes, while more significant changes were noted in sensitivity to nucleoside 5'-triphosphate analogues that inhibit the enzyme activity. Results suggest that altered substrate recognition by the HIV-1 RT is involved in the mechanism of resistance.Keywords
This publication has 22 references indexed in Scilit:
- Use of evolutionary limitations of HIV-1 multidrug resistance to optimize therapyNature, 1993
- Multiple Mutations in HIV-1 Reverse Transcriptase Confer High-Level Resistance to Zidovudine (AZT)Science, 1989
- Studies on the mechanism of human immunodeficiency virus reverse transcriptase. Steady-state kinetics, processivity, and polynucleotide inhibition.Journal of Biological Chemistry, 1988
- 3'-Azido-3'-deoxythymidine triphosphate as an inhibitor and substrate of purified human immunodeficiency virus reverse transcriptaseAntimicrobial Agents and Chemotherapy, 1987
- Phosphorylation of 3'-azido-3'-deoxythymidine and selective interaction of the 5'-triphosphate with human immunodeficiency virus reverse transcriptase.Proceedings of the National Academy of Sciences, 1986
- Characterization of Highly Immunogenic p66/p51 as the Reverse Transcriptase of HTLV-III/LAVScience, 1986
- Characterization of the AIDS-associated retrovirus reverse transcriptase and optimal conditions for its detection in virionsVirology, 1985
- A molecular clone of HTLV-III with biological activityNature, 1985
- Characterization of the RNA dependent DNA polymerase of a new human T lymphotropic retrovirus (lymphadenopathy associated virus)Biochemical and Biophysical Research Communications, 1984
- Elementary steps in the DNA polymerase I reaction pathwayBiochemistry, 1983