title: Mutations E44D and V118I in the reverse transcriptase of HIV-1 play distinct mechanistic roles in dual resistance to AZT and 3TC.
abstract: Retroviral resistance to AZT and 3TC has been associated with two different mechanisms. The M184V mutation in the reverse transcriptase (RT) of the human immunodeficiency virus, type 1 (HIV-1) diminishes the incorporation of 3TC-monophosphate (3TC-MP), whereas AZT resistance-conferring mutations were shown to facilitate the phosphorolytic excision of incorporated AZT-MP in the presence of ATP. Both mechanisms show a certain degree of incompatibility; however, previous clinical data revealed that mutations E44D and V118I, when present in a background of classical AZT mutations (M41L, D67N, L210W, and T215Y), confer dual resistance to AZT and 3TC. We have purified RT enzymes that contain E44D and V118I either alone or in a background of different combinations of AZT mutations to study the underlying biochemical mechanisms. We found that enzymes containing E44D in a background of these latter mutations increase the efficiency of excision of 3TC-MP. Unexpectedly, V118I-containing enzymes show dramatic reductions in rates of incorporation of AZT-MP and 3TC-MP. The V118I mutant is also associated with diminished rates of ATP-dependent primer unblocking. The additional presence of mutations M41L, D67N, L210W, and T215Y can partially neutralize this deficit, which helps to explain the concurrent presence of these changes in resistant isolates. These biochemical data make clear that mutations E44D and V118I play distinct mechanistic roles in dual resistance to AZT and 3TC. Our findings are consistent with an increasing number of clinical studies suggesting that the V118I cluster constitutes a novel pathway for HIV resistance to multiple nucleotide analogue RT inhibitors.
authors.affiliation: McGill University AIDS Centre, Lady Davis Institute-Jewish General Hospital and Department of Microbiology and Immunology, McGill University, Montréal, Québec, Canada.
authors.forename: Mélanie
authors.initials: M
authors.lastname: Girouard
authors.forename: Karidia
authors.initials: K
authors.lastname: Diallo
authors.forename: Bruno
authors.initials: B
authors.lastname: Marchand
authors.forename: Suzanne
authors.initials: S
authors.lastname: McCormick
authors.forename: Matthias
authors.initials: M
authors.lastname: Götte
mesh_terms: D000255:Adenosine Triphosphate / Q000737:chemistry; D019380:Anti-HIV Agents / Q000494:pharmacology*; D001483:Base Sequence; D004247:DNA / Q000378:metabolism; D004252:DNA Mutational Analysis; D004305:Dose-Response Relationship, Drug; D024882:Drug Resistance, Viral; D054303:HIV Reverse Transcriptase / Q000737:chemistry / Q000235:genetics*; D020128:Inhibitory Concentration 50; D007700:Kinetics; D019259:Lamivudine / Q000494:pharmacology*; D008969:Molecular Sequence Data; D009154:Mutation; D010641:Phenotype; D013997:Time Factors; D015215:Zidovudine / Q000494:pharmacology*
chemical_list: D019380:Anti-HIV Agents; D019259:Lamivudine; D015215:Zidovudine; D000255:Adenosine Triphosphate; D004247:DNA; D054303:HIV Reverse Transcriptase