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

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 withmore » therapeutics.« less
Authors:
 [1] ;  [1] ;  [1] ;  [1] ;  [1] ;  [1]
  1. National Institute of Environmental Health Sciences, Research Triangle Park, NC (United States)
Publication Date:
Grant/Contract Number:
W-31-109-ENG-38; AC02-06CH11357
Type:
Accepted Manuscript
Journal Name:
Nucleic Acids Research
Additional Journal Information:
Journal Volume: 44; Journal Issue: 4; Journal ID: ISSN 0305-1048
Publisher:
Oxford University Press
Research Org:
Argonne National Lab. (ANL), Argonne, IL (United States)
Sponsoring Org:
USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22)
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES
OSTI Identifier:
1251217

Zheng, Xunhai, Pedersen, Lars C., Gabel, Scott A., Mueller, Geoffrey A., DeRose, Eugene F., and London, Robert E.. Unfolding the HIV-1 reverse transcriptase RNase H domain – how to lose a molecular tug-of-war. United States: N. p., Web. doi:10.1093/nar/gkv1538.
Zheng, Xunhai, Pedersen, Lars C., Gabel, Scott A., Mueller, Geoffrey A., DeRose, Eugene F., & London, Robert E.. Unfolding the HIV-1 reverse transcriptase RNase H domain – how to lose a molecular tug-of-war. United States. doi:10.1093/nar/gkv1538.
Zheng, Xunhai, Pedersen, Lars C., Gabel, Scott A., Mueller, Geoffrey A., DeRose, Eugene F., and London, Robert E.. 2016. "Unfolding the HIV-1 reverse transcriptase RNase H domain – how to lose a molecular tug-of-war". United States. doi:10.1093/nar/gkv1538. https://www.osti.gov/servlets/purl/1251217.
@article{osti_1251217,
title = {Unfolding the HIV-1 reverse transcriptase RNase H domain – how to lose a molecular tug-of-war},
author = {Zheng, Xunhai and Pedersen, Lars C. and Gabel, Scott A. and Mueller, Geoffrey A. and DeRose, Eugene F. and London, Robert E.},
abstractNote = {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.},
doi = {10.1093/nar/gkv1538},
journal = {Nucleic Acids Research},
number = 4,
volume = 44,
place = {United States},
year = {2016},
month = {1}
}