Unfolding the HIV-1 reverse transcriptase RNase H domain – how to lose a molecular tug-of-war
Abstract
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 »
- Authors:
-
- National Institute of Environmental Health Sciences, Research Triangle Park, NC (United States)
- Publication Date:
- Research Org.:
- Argonne National Laboratory (ANL), Argonne, IL (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC), Basic Energy Sciences (BES)
- OSTI Identifier:
- 1251217
- Grant/Contract Number:
- W-31-109-ENG-38; AC02-06CH11357
- Resource Type:
- Journal Article: 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
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES
Citation Formats
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., 2016.
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. https://doi.org/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. https://doi.org/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},
url = {https://www.osti.gov/biblio/1251217},
journal = {Nucleic Acids Research},
issn = {0305-1048},
number = 4,
volume = 44,
place = {United States},
year = {Thu Jan 14 00:00:00 EST 2016},
month = {Thu Jan 14 00:00:00 EST 2016}
}
Web of Science
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