Structural basis of deoxynucleotide addition by HIV-1 RT during reverse transcription.

ABSTRACT: Reverse transcription of the retroviral RNA genome into DNA is an integral step during HIV-1 replication. Despite a wealth of structural information on reverse transcriptase (RT), we lack insight into the intermediate states of DNA synthesis. Using catalytically active substrates, and a blot/diffusion cryo-electron microscopy approach, we capture 11 structures encompassing reactant, intermediate and product states of dATP addition by RT at 2.2 to 3.0 Å resolution. In the reactant state, dATP binding to RT-template/primer involves a single Mg²⁺ (site B) inducing formation of a negatively charged pocket where a second floating Mg²⁺ can bind (site A). During the intermediate state, the α-phosphate oxygen from a previously unobserved dATP conformer aligns with site A Mg

SUBMITTER: Vergara S 

PROVIDER: S-EPMC11618517 | biostudies-literature | 2024 Dec

REPOSITORIES: biostudies-literature