Year: 2020
Type: article
Abstract: BACKGROUND: Antiretroviral therapy (ART) can mitigate the morbidity and mortality caused by the human immunodeficiency virus (HIV). Successful development of ART can be accelerated by accurate structural and biochemical data on targets and their responses to inhibitors. One important ART target, HIV integrase (IN), has historically been studied in vitro in a modified form adapted to bacterial overexpression, with a methionine or a longer fusion protein sequence at the N-terminus. In contrast, IN present in viral particles is produced by proteolytic cleavage of the Pol polyprotein, which leaves a phenylalanine at the N-terminus (IN 1F). Inspection of available structures suggested that added residues on the N-terminus might disrupt proper protein folding and formation of multimeric complexes. RESULTS: . Full-length IN 1F showed increased in vitro catalytic activity in assays of coupled joining of the two viral DNA ends compared to two IN variants containing additional N-terminal residues. IN 1F was also altered in its sensitivity to inhibitors, showing decreased sensitivity to the strand-transfer inhibitor raltegravir and increased sensitivity to allosteric integrase inhibitors. In solution, IN 1F exists as monomers and dimers, in contrast to other IN preparations which exist as higher-order oligomers. CONCLUSIONS: The structural, biochemical, and biophysical characterization of IN 1F reveals the conformation of the native HIV-1 IN N-terminus and accompanying unique biochemical and biophysical properties. IN 1F thus represents an improved reagent for use in integration reactions in vitro and the development of antiretroviral agents.
Source: Retrovirology
Authors: Grant Eilers, Kushol Gupta, Audrey Allen, Jeffrey Zhou, Young Hwang, Michael B. Cory, Frederic D. Bushman, Gregory Van Duyne
Institution: University of Pennsylvania
Language: English
Subfield: Infectious Diseases
Field: Medicine
Domain: Health Sciences
Open Access status: gold
Estimated APC paid: $2,790
Funders: U.S. Department of Energy, University of Pennsylvania, Office of Science, Center for AIDS Research, University of Washington, National Heart, Lung, and Blood Institute, National Institute of Allergy and Infectious Diseases, National Institute of Neurological Disorders and Stroke, Brookhaven National Laboratory
Awards: BEAT-HIV: Delaney Collaboratory to Cure HIV-1 Infection by Combination Immunotherapy, A New View of PAH Allostery - Correlation with Disease-Associated Alleles, The Lung DNA Virome in Health and Disease, HIV Pathogenesis, Vaccination and Cure, Optimization HIV Inhibition by Allosteric Integrase Inhibitors, Engineering T cells to Provide Durable Control of HIV-1 Replication, Lung Transplant Microbiome and Chronic Allograft Dysfunction, Penn Center for AIDS Research
OpenAlex ID: W3081563294