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Upstream of N-Ras (Unr/CSDE1) Interacts with NCp7 and Gag, Modulating HIV-1 IRES-Mediated Translation Initiation

The Human Immunodeficiency Virus-1 (HIV-1) nucleocapsid protein (NC) as a mature protein or as a domain of the Gag precursor plays important roles in the early and late phases of the infection. To better understand its roles, we searched for new cellular partners and identified the RNA-binding prote...

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Autores principales: Taha, Nedal, Zgheib, Sarwat, Sharma, Kamal Kant, Humbert, Nicolas, Boutant, Emmanuel, Didier, Pascal, Mély, Yves, Real, Eleonore
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9413769/
https://www.ncbi.nlm.nih.gov/pubmed/36016420
http://dx.doi.org/10.3390/v14081798
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author Taha, Nedal
Zgheib, Sarwat
Sharma, Kamal Kant
Humbert, Nicolas
Boutant, Emmanuel
Didier, Pascal
Mély, Yves
Real, Eleonore
author_facet Taha, Nedal
Zgheib, Sarwat
Sharma, Kamal Kant
Humbert, Nicolas
Boutant, Emmanuel
Didier, Pascal
Mély, Yves
Real, Eleonore
author_sort Taha, Nedal
collection PubMed
description The Human Immunodeficiency Virus-1 (HIV-1) nucleocapsid protein (NC) as a mature protein or as a domain of the Gag precursor plays important roles in the early and late phases of the infection. To better understand its roles, we searched for new cellular partners and identified the RNA-binding protein Unr/CSDE1, Upstream of N-ras, whose interaction with Gag and NCp7 was confirmed by co-immunoprecipitation and FRET-FLIM. Unr interaction with Gag was found to be RNA-dependent and mediated by its NC domain. Using a dual luciferase assay, Unr was shown to act as an ITAF (IRES trans-acting factor), increasing the HIV-1 IRES-dependent translation. Point mutations of the HIV-1 IRES in a consensus Unr binding motif were found to alter both the IRES activity and its activation by Unr, suggesting a strong dependence of the IRES on Unr. Interestingly, Unr stimulatory effect is counteracted by NCp7, while Gag increases the Unr-promoted IRES activity, suggesting a differential Unr effect on the early and late phases of viral infection. Finally, knockdown of Unr in HeLa cells leads to a decrease in infection by a non-replicative lentivector, proving its functional implication in the early phase of viral infection.
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spelling pubmed-94137692022-08-27 Upstream of N-Ras (Unr/CSDE1) Interacts with NCp7 and Gag, Modulating HIV-1 IRES-Mediated Translation Initiation Taha, Nedal Zgheib, Sarwat Sharma, Kamal Kant Humbert, Nicolas Boutant, Emmanuel Didier, Pascal Mély, Yves Real, Eleonore Viruses Article The Human Immunodeficiency Virus-1 (HIV-1) nucleocapsid protein (NC) as a mature protein or as a domain of the Gag precursor plays important roles in the early and late phases of the infection. To better understand its roles, we searched for new cellular partners and identified the RNA-binding protein Unr/CSDE1, Upstream of N-ras, whose interaction with Gag and NCp7 was confirmed by co-immunoprecipitation and FRET-FLIM. Unr interaction with Gag was found to be RNA-dependent and mediated by its NC domain. Using a dual luciferase assay, Unr was shown to act as an ITAF (IRES trans-acting factor), increasing the HIV-1 IRES-dependent translation. Point mutations of the HIV-1 IRES in a consensus Unr binding motif were found to alter both the IRES activity and its activation by Unr, suggesting a strong dependence of the IRES on Unr. Interestingly, Unr stimulatory effect is counteracted by NCp7, while Gag increases the Unr-promoted IRES activity, suggesting a differential Unr effect on the early and late phases of viral infection. Finally, knockdown of Unr in HeLa cells leads to a decrease in infection by a non-replicative lentivector, proving its functional implication in the early phase of viral infection. MDPI 2022-08-17 /pmc/articles/PMC9413769/ /pubmed/36016420 http://dx.doi.org/10.3390/v14081798 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/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 (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Taha, Nedal
Zgheib, Sarwat
Sharma, Kamal Kant
Humbert, Nicolas
Boutant, Emmanuel
Didier, Pascal
Mély, Yves
Real, Eleonore
Upstream of N-Ras (Unr/CSDE1) Interacts with NCp7 and Gag, Modulating HIV-1 IRES-Mediated Translation Initiation
title Upstream of N-Ras (Unr/CSDE1) Interacts with NCp7 and Gag, Modulating HIV-1 IRES-Mediated Translation Initiation
title_full Upstream of N-Ras (Unr/CSDE1) Interacts with NCp7 and Gag, Modulating HIV-1 IRES-Mediated Translation Initiation
title_fullStr Upstream of N-Ras (Unr/CSDE1) Interacts with NCp7 and Gag, Modulating HIV-1 IRES-Mediated Translation Initiation
title_full_unstemmed Upstream of N-Ras (Unr/CSDE1) Interacts with NCp7 and Gag, Modulating HIV-1 IRES-Mediated Translation Initiation
title_short Upstream of N-Ras (Unr/CSDE1) Interacts with NCp7 and Gag, Modulating HIV-1 IRES-Mediated Translation Initiation
title_sort upstream of n-ras (unr/csde1) interacts with ncp7 and gag, modulating hiv-1 ires-mediated translation initiation
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9413769/
https://www.ncbi.nlm.nih.gov/pubmed/36016420
http://dx.doi.org/10.3390/v14081798
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