Biochemistry (Easton), 1996-02, Vol.35 (6), p.1911-1917
1996
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- Autor(en) / Beteiligte
- Titel
- Characterization of the p68/p58 Heterodimer of Human Immunodeficiency Virus Type 2 Reverse Transcriptase
- Ist Teil von
- Biochemistry (Easton), 1996-02, Vol.35 (6), p.1911-1917
- Ort / Verlag
- United States: American Chemical Society
- Erscheinungsjahr
- 1996
- Quelle
- MEDLINE
- Beschreibungen/Notizen
- Recently we demonstrated that the p58 subunit of p68/p58 HIV-2 reverse transcriptase (RT) heterodimer, produced by processing of p68/p68 homodimer with recombinant HIV-2 protease, terminates at Met484 [Fan, N., et al. (1995) J. Biol. Chem. 270, 13573−13579]. Here we describe purification and characterization of the p68/p58 heterodimer of recombinant HIV-2 RT. It exhibited both RT and RNase H activities, obeyed Michaelis−Menten kinetics, and was competitively inhibited by the DNA chain terminator ddTTP (K i[app] = 305 ± 20 nM). The HIV-2 RT-associated RNase H exhibited a marked preference for RNA hydrolysis from a HIV-1 gag-based heteropolymeric RNA/DNA hybrid in the presence of either Mg2+ or Mn2+, compared to the [3H]poly(rA)·poly(dT) or [3H]poly(rG)·poly(dC) homopolymeric substrates. Relative to HIV-1 RT, the RNase H activity of HIV-2 RT was only 5% toward the [3H]poly(rA)·poly(dT) in the presence of Mg2+. The size distribution of products generated from [3H]poly(rA)· poly(dT) by HIV-2 RT-associated RNase H was markedly distinct from that of HIV-1 RT in the presence of Mg2+ or Mn2+. The p68/p58 HIV-2 RT heterodimer, produced by specific cleavage using HIV-2 protease, should be useful for inhibition and biophysical studies aimed at discovering and designing drugs directed toward HIV-2.
- Sprache
- Englisch
- Identifikatoren
- ISSN: 0006-2960eISSN: 1520-4995DOI: 10.1021/bi9516440Titel-ID: cdi_proquest_miscellaneous_78043039
- Format
- –
- Schlagworte
- AIDS/HIV, Aspartic Acid Endopeptidases - metabolism, Base Sequence, Cations, Divalent - pharmacology, Dideoxynucleotides, DNA Primers - genetics, DNA, Viral - genetics, Genes, gag, HIV Protease, HIV Reverse Transcriptase, HIV-2 - enzymology, HIV-2 - genetics, human immunodeficiency virus 2, Humans, In Vitro Techniques, Kinetics, Molecular Sequence Data, Poly A - metabolism, Poly T - metabolism, Protein Conformation, Protein Processing, Post-Translational, Recombinant Proteins - chemistry, Recombinant Proteins - genetics, Recombinant Proteins - metabolism, Reverse Transcriptase Inhibitors - pharmacology, Ribonuclease H - metabolism, RNA-Directed DNA Polymerase - chemistry, RNA-Directed DNA Polymerase - genetics, RNA-Directed DNA Polymerase - metabolism, Substrate Specificity, Thymine Nucleotides - pharmacology