Unfolding the HIV-1 reverse transcriptase RNase H domain – how to lose a molecular tug-of-war

Zheng, Xunhai; Pedersen, Lars C.; Gabel, Scott A.; Mueller, Geoffrey A.; DeRose, Eugene F.; London, Robert E.

doi:10.1093/nar/gkv1538

Title: Unfolding the HIV-1 reverse transcriptase RNase H domain – how to lose a molecular tug-of-war

Journal Article · Thu Jan 14 00:00:00 EST 2016 · Nucleic Acids Research

DOI:https://doi.org/10.1093/nar/gkv1538· OSTI ID:1251217

Zheng, Xunhai ^[1]; Pedersen, Lars C. ^[1]; Gabel, Scott A. ^[1]; Mueller, Geoffrey A. ^[1]; DeRose, Eugene F. ^[1]; London, Robert E. ^[1]

National Institute of Environmental Health Sciences, Research Triangle Park, NC (United States)

Formation of the mature HIV-1 reverse transcriptase (RT) p66/p51 heterodimer requires subunit-specific processing of the p66/p66' homodimer precursor. Since the ribonuclease H (RH) domain contains an occult cleavage site located near its center, cleavage must occur either prior to folding or subsequent to unfolding. Recent NMR studies have identified a slow, subunit-specific RH domain unfolding process proposed to result from a residue tug-of-war between the polymerase and RH domains on the functionally inactive, p66' subunit. Here, we describe a structural comparison of the isolated RH domain with a domain swapped RH dimer that reveals several intrinsically destabilizing characteristics of the isolated domain that facilitate excursions of Tyr427 from its binding pocket and separation of helices B and D. These studies provide independent support for the subunit-selective RH domain unfolding pathway in which instability of the Tyr427 binding pocket facilitates its release followed by domain transfer, acting as a trigger for further RH domain destabilization and subsequent unfolding. As further support for this pathway, NMR studies demonstrate that addition of an RH active site-directed isoquinolone ligand retards the subunit-selective RH' domain unfolding behavior of the p66/p66' homodimer. As a result, this study demonstrates the feasibility of directly targeting RT maturation with therapeutics.

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Research Organization:: Argonne National Laboratory (ANL), Argonne, IL (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Basic Energy Sciences (BES)

Grant/Contract Number:: W-31-109-ENG-38; AC02-06CH11357

OSTI ID:: 1251217

Journal Information:: Nucleic Acids Research, Vol. 44, Issue 4; ISSN 0305-1048

Publisher:: Oxford University PressCopyright Statement

Country of Publication:: United States

Language:: English

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Cited by: 11 works

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