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Functional Equivalence of Retroviral MA Domains in Facilitating Psi RNA Binding Specificity by Gag

Retroviruses specifically package full-length, dimeric genomic RNA (gRNA) even in the presence of a vast excess of cellular RNA. The “psi” (Ψ) element within the 5′-untranslated region (5′UTR) of gRNA is critical for packaging through interaction with the nucleocapsid (NC) domain of Gag. However, in...

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Autores principales: Rye-McCurdy, Tiffiny, Olson, Erik D., Liu, Shuohui, Binkley, Christiana, Reyes, Joshua-Paolo, Thompson, Brian R., Flanagan, John M., Parent, Leslie J., Musier-Forsyth, Karin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5035970/
https://www.ncbi.nlm.nih.gov/pubmed/27657107
http://dx.doi.org/10.3390/v8090256
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author Rye-McCurdy, Tiffiny
Olson, Erik D.
Liu, Shuohui
Binkley, Christiana
Reyes, Joshua-Paolo
Thompson, Brian R.
Flanagan, John M.
Parent, Leslie J.
Musier-Forsyth, Karin
author_facet Rye-McCurdy, Tiffiny
Olson, Erik D.
Liu, Shuohui
Binkley, Christiana
Reyes, Joshua-Paolo
Thompson, Brian R.
Flanagan, John M.
Parent, Leslie J.
Musier-Forsyth, Karin
author_sort Rye-McCurdy, Tiffiny
collection PubMed
description Retroviruses specifically package full-length, dimeric genomic RNA (gRNA) even in the presence of a vast excess of cellular RNA. The “psi” (Ψ) element within the 5′-untranslated region (5′UTR) of gRNA is critical for packaging through interaction with the nucleocapsid (NC) domain of Gag. However, in vitro Gag binding affinity for Ψ versus non-Ψ RNAs is not significantly different. Previous salt-titration binding assays revealed that human immunodeficiency virus type 1 (HIV-1) Gag bound to Ψ RNA with high specificity and relatively few charge interactions, whereas binding to non-Ψ RNA was less specific and involved more electrostatic interactions. The NC domain was critical for specific Ψ binding, but surprisingly, a Gag mutant lacking the matrix (MA) domain was less effective at discriminating Ψ from non-Ψ RNA. We now find that Rous sarcoma virus (RSV) Gag also effectively discriminates RSV Ψ from non-Ψ RNA in a MA-dependent manner. Interestingly, Gag chimeras, wherein the HIV-1 and RSV MA domains were swapped, maintained high binding specificity to cognate Ψ RNAs. Using Ψ RNA mutant constructs, determinants responsible for promoting high Gag binding specificity were identified in both systems. Taken together, these studies reveal the functional equivalence of HIV-1 and RSV MA domains in facilitating Ψ RNA selectivity by Gag, as well as Ψ elements that promote this selectivity.
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spelling pubmed-50359702016-09-29 Functional Equivalence of Retroviral MA Domains in Facilitating Psi RNA Binding Specificity by Gag Rye-McCurdy, Tiffiny Olson, Erik D. Liu, Shuohui Binkley, Christiana Reyes, Joshua-Paolo Thompson, Brian R. Flanagan, John M. Parent, Leslie J. Musier-Forsyth, Karin Viruses Article Retroviruses specifically package full-length, dimeric genomic RNA (gRNA) even in the presence of a vast excess of cellular RNA. The “psi” (Ψ) element within the 5′-untranslated region (5′UTR) of gRNA is critical for packaging through interaction with the nucleocapsid (NC) domain of Gag. However, in vitro Gag binding affinity for Ψ versus non-Ψ RNAs is not significantly different. Previous salt-titration binding assays revealed that human immunodeficiency virus type 1 (HIV-1) Gag bound to Ψ RNA with high specificity and relatively few charge interactions, whereas binding to non-Ψ RNA was less specific and involved more electrostatic interactions. The NC domain was critical for specific Ψ binding, but surprisingly, a Gag mutant lacking the matrix (MA) domain was less effective at discriminating Ψ from non-Ψ RNA. We now find that Rous sarcoma virus (RSV) Gag also effectively discriminates RSV Ψ from non-Ψ RNA in a MA-dependent manner. Interestingly, Gag chimeras, wherein the HIV-1 and RSV MA domains were swapped, maintained high binding specificity to cognate Ψ RNAs. Using Ψ RNA mutant constructs, determinants responsible for promoting high Gag binding specificity were identified in both systems. Taken together, these studies reveal the functional equivalence of HIV-1 and RSV MA domains in facilitating Ψ RNA selectivity by Gag, as well as Ψ elements that promote this selectivity. MDPI 2016-09-19 /pmc/articles/PMC5035970/ /pubmed/27657107 http://dx.doi.org/10.3390/v8090256 Text en © 2016 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC-BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Rye-McCurdy, Tiffiny
Olson, Erik D.
Liu, Shuohui
Binkley, Christiana
Reyes, Joshua-Paolo
Thompson, Brian R.
Flanagan, John M.
Parent, Leslie J.
Musier-Forsyth, Karin
Functional Equivalence of Retroviral MA Domains in Facilitating Psi RNA Binding Specificity by Gag
title Functional Equivalence of Retroviral MA Domains in Facilitating Psi RNA Binding Specificity by Gag
title_full Functional Equivalence of Retroviral MA Domains in Facilitating Psi RNA Binding Specificity by Gag
title_fullStr Functional Equivalence of Retroviral MA Domains in Facilitating Psi RNA Binding Specificity by Gag
title_full_unstemmed Functional Equivalence of Retroviral MA Domains in Facilitating Psi RNA Binding Specificity by Gag
title_short Functional Equivalence of Retroviral MA Domains in Facilitating Psi RNA Binding Specificity by Gag
title_sort functional equivalence of retroviral ma domains in facilitating psi rna binding specificity by gag
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5035970/
https://www.ncbi.nlm.nih.gov/pubmed/27657107
http://dx.doi.org/10.3390/v8090256
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