Effect of tRNA on the Maturation of HIV-1 Reverse Transcriptase

• Type of Material: Article
• Authors: Tatiana V. Ilina, University of Pittsburgh; Ryan L. Slack, University of Pittsburgh; John H. Elder, Scripps Research Institute; Stefan Sarafianos, Emory University; Michael A. Parniak, University of Pittsburgh; Rieko Ishima, University of Pittsburgh
• Language: English
• Date: 2018-06-22
• Publisher: Elsevier
• Publication Version: Author Accepted Manuscript (After Peer Review)
• Copyright Statement: © 2018 Elsevier Ltd
• License: Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International
• Final Published Version (URL): http://dx.doi.org/10.1016/j.jmb.2018.02.027
• Title of Journal or Parent Work: Journal of Molecular Biology
• ISSN: 0022-2836
• Volume: 430
• Issue: 13
• Start Page: 1891
• End Page: 1900
• Grant/Funding Information: This study was supported by grants from the National Institutes of Health (R01GM105401 to RI and RLS, R01GM109767 to RI, R01AI00890 to TI, SGS, and MAP, P50GM103368 to JHE, SGS, and MAP).
• Keywords: reverse transcriptase; RNase H; HIV-1; CRYSTAL-STRUCTURE; POL POLYPROTEIN PRECURSOR; RIBONUCLEASE-H DOMAIN; IN-VITRO; Life Sciences & Biomedicine; PROTEASE INHIBITORS; Science & Technology; HOMODIMER FORMATION; proteolysis; maturation; Biochemistry & Molecular Biology; tRNA; IMMUNODEFICIENCY-VIRUS TYPE-1; ANGSTROM RESOLUTION; GAG-POL; TRYPTOPHAN REPEAT MOTIF
• Research Categories: Biology, Molecular; Chemistry, Biochemistry; Health Sciences, Public Health

Abstract

The mature HIV-1 reverse transcriptase is a heterodimer that comprises 66 kDa (p66) and 51 kDa (p51) subunits. The latter is formed by HIV-1 protease-catalyzed removal of a C-terminal ribonuclease H domain from a p66 subunit. This proteolytic processing is a critical step in virus maturation and essential for viral infectivity. Here, we report that tRNA significantly enhances in vitro processing even at a substoichiometric tRNA:p66/p66 ratio. Other double-stranded RNAs have considerably less pronounced effect. Our data support a model where interaction of p66/p66 with tRNA introduces conformational asymmetry in the two subunits, permitting specific proteolytic processing of one p66 to provide the mature RT p66/p51 heterodimer.