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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...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2016
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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. |
format | Online Article Text |
id | pubmed-5035970 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
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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