title: The impact of molecular manipulation in residue 114 of human immunodeficiency virus type-1 reverse transcriptase on dNTP substrate binding and viral replication.
abstract: Human immunodeficiency virus type-1 (HIV-1) reverse transcriptase (RT) has a unique tight binding to dNTP substrates. Structural modeling of Ala-114 of HIV-1 RT suggests that longer side chains at this residue can reduce the space normally occupied by the sugar moiety of an incoming dNTP. Indeed, mutations at Ala-114 decrease the ability of RT to synthesize DNA at low dNTP concentrations and reduce the dNTP-binding affinity (K(d)) of RT. However, the K(d) values of WT and A114C RT remained equivalent with an acyclic dNTP substrate. Finally, mutant A114 RT HIV-1 vectors displayed a greatly reduced transduction in nondividing human lung fibroblasts (HLFs), while WT HIV-1 vector efficiently transduced both dividing and nondividing HLFs. Together these data support that the A114 residue of HIV-1 RT plays a key mechanistic role in the dNTP binding of HIV-1 RT and the unique viral infectivity of target cell types with low dNTP pools.
authors.affiliation: Department of Microbiology and Immunology, University of Rochester Medical Center, Rochester, NY 14642, USA.
authors.forename: Sarah K
authors.initials: SK
authors.lastname: Van Cor-Hosmer
authors.forename: Waaqo
authors.initials: W
authors.lastname: Daddacha
authors.forename: Z
authors.initials: Z
authors.lastname: Kelly
authors.forename: Amy
authors.initials: A
authors.lastname: Tsurumi
authors.forename: Edward M
authors.initials: EM
authors.lastname: Kennedy
authors.forename: Baek
authors.initials: B
authors.lastname: Kim
mesh_terms: D001665:Binding Sites; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology*; D015967:Gene Expression Regulation, Viral / Q000502:physiology; D054303:HIV Reverse Transcriptase / Q000235:genetics / Q000378:metabolism*; D015497:HIV-1 / Q000201:enzymology* / Q000502:physiology*; D006801:Humans; D007700:Kinetics; D008958:Models, Molecular; D009705:Nucleosides / Q000378:metabolism*; D011485:Protein Binding; D013379:Substrate Specificity; D014779:Virus Replication / Q000502:physiology*
chemical_list: D009705:Nucleosides; C514824:reverse transcriptase, Human immunodeficiency virus 1; D054303:HIV Reverse Transcriptase