Conformational Changes in HIV-1 Reverse Transcriptase that Facilitate Its Maturation

• Type of Material: Article
• Authors: Ryan L. Slack, University of Pittsburgh; Tatiana V. Ilina, University of Pittsburgh; Zhaoyong Xi, University of Pittsburgh; Nicholas S. Giacobbi, University of Pittsburgh; Gota Kawai, Chiba Institute of Technology; Michael A. Parniak, University of Pittsburgh; Stefan Sarafianos, Emory University; Nicolas Sluis Cremer, University of Pittsburgh; Rieko Ishima, University of Pittsburgh
• Language: English
• Date: 2019-10-01
• Publisher: CELL PRESS
• Publication Version: Author Accepted Manuscript (After Peer Review)
• Copyright Statement: © 2019 Elsevier Ltd.
• License: Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International
• Final Published Version (URL): http://dx.doi.org/10.1016/j.str.2019.08.004
• Title of Journal or Parent Work: STRUCTURE
• Volume: 27
• Issue: 10
• Start Page: 1581
• End Page: +
• Grant/Funding Information: This study was supported by grants from the National Institutes of Health (R01GM105401 to R.I., U54AI150472 and R01GM118012 to S.G.S, R01GM068406 to N.S.C, and P50AI150481 to R.I. and N.S.C.), and the MEXT-Supported Program for the Strategic Research Foundation at Private Universities in Japan (the term, 2011-2015, to G.K.).
• Keywords: POL POLYPROTEIN PRECURSOR; EFAVIRENZ BINDING; RIBONUCLEASE-H DOMAIN; Cell Biology; IMMUNODEFICIENCY-VIRUS TYPE-1; Biophysics; CRYSTAL-STRUCTURE; WILD-TYPE; TRNA(LYS) INCORPORATION; Biochemistry & Molecular Biology; Life Sciences & Biomedicine; Science & Technology; ANGSTROM RESOLUTION; TRANSFER-RNA SYNTHETASE; GAGPOL PRECURSOR

Abstract

HIV-1 reverse transcriptase (RT) is translated as part of the Gag-Pol polyprotein that is proteolytically processed by HIV-1 protease (PR) to finally become a mature heterodimer, composed of a p66 and a p66-derived 51-kDa subunit, p51. Our previous work suggested that tRNALys3 binding to p66/p66 introduces conformational changes in the ribonuclease (RNH) domain of RT that facilitate efficient cleavage of p66 to p51 by PR. In this study, we characterized the conformational changes in the RNH domain of p66/p66 imparted by tRNALys3 using NMR. Moreover, the importance of tRNALys3 in RT maturation was confirmed in cellulo by modulating the levels of Lys-tRNA synthetase, which affects recruitment of tRNALys3 to the virus. We also employed nonnucleoside RT inhibitors, to modulate the p66 dimer-monomer equilibrium and monitor the resulting structural changes. Taken together, our data provide unique insights into the conformational changes in p66/p66 that drive PR cleavage.