A Thermodynamic Analysis of the Binding of Nucleic Acid to HIV-1 Reverse Transcriptase
Human immunodeficiency virus type 1 (HIV-1) encodes a Mg+2-dependent reverse transcriptase (E.C.220.127.116.11) that synthesizes a double-stranded DNA copy of genomic RNA. The enzyme purified from virions has been shown to consist of two polypeptides of molecular weights 66,000 and 51,000 (Hoffman et al., 1985; Di Marzo Veronese et al.,1986). The 66 kD subunit has both polymerase and RNase H activities. Like other retroviral reverse transcriptases, the associated RNase H activity is located on the carboxy terminal portion of the polypeptide (Johnson et al., 1986; Hansen et al., 1987; Tisdale et al., 1988). The 51 kD subunit is derived from the 66 kD polypeptide by cleavage at a protease sensitive site on the linker between the polymerase and RNase H domains (Lowe et al.,1988). Catalytically active HIV-1 reverse transcriptase has been cloned and expressed in E. coli (Larder et al., 1987). The recombinant, heterodimeric enzyme is kinetically indistinguishable from the native enzyme purified from virus.
KeywordsBinary Complex Klenow Fragment Polymerase Domain Human Immunodeficiency Virus Reverse Transcriptase Heterodimeric Enzyme
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- Helene, C., 1971, Role of aromatic amino acid residues in the binding of enzymes and proteins to nucleic acids, Nature (London), New Biol., 234 (47): 120–121.Google Scholar
- Kornberg, A., 1974, “DNA Synthesis,” W.H. Freeman and Co., San Francisco.Google Scholar
- Lakowicz, J.R., 1983, “Principles of Fluorescence Spectroscopy,” Plenum Press, New York.Google Scholar
- Steitz, T.A., Weber, I.T., and Matthew, J.B., 1983, Catabolite gene activator protein: Structure, homology with other proteins, and cyclic AMP and DNA binding, Cold Spring Harbor Symp. Quant. Biol. 1982, 47 (1): 419–426.Google Scholar
- Still, W.C., Richards, N.G., Guida, W.C., Lipton, M., Liskamp, R., Chang, G., and Hendrickson, T. 1989, MacroModel Version 2. 5, Dept. of Chemistry, Columbia University.Google Scholar
- Thomas, D., Griffith, J., Furman, P., and Painter, G., 1990, Electron microscopic visualization of HIV-1 reverse transcriptase free and bound to DNA, J. Cellular Biology, 14D: 112.Google Scholar
- Tisdale, M., Ertl, P., Larder, B.A., Purifoy, D.J.M., Darby, G.K., and Powell, K.L., 1988, Characterization of human immunodeficiency virus type 1 reverse transcriptase by using monoclonal antibodies: Role of the C terminus in antibody reactivity and enzyme function, J. Virol., 62: 3662–3667.PubMedGoogle Scholar