C. Webster Andrews

Conformation and Local Environment of Nucleotides Bound To Hiv Type 1 Reverse Transcriptase (HIV-1 RT) in The Ground State

Abstract Nuclear Overhauser effect experiments were used to characterize the protein environment and conformations of dTTP, dATP and AZTTP bound to HIV-RT in the ground state. The results show the initial binding sites for the nucleotides overlap but are not completely coincident. All of the bound nucleotides assume the same anti C4′-exo conformation.

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.2.7.7.49) 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.