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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MDPI AG
2022-08-01
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| Online Access: | https://www.mdpi.com/1999-4915/14/8/1798 |
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| author | Nedal Taha Sarwat Zgheib Kamal Kant Sharma Nicolas Humbert Emmanuel Boutant Pascal Didier Yves Mély Eleonore Real |
| author_facet | Nedal Taha Sarwat Zgheib Kamal Kant Sharma Nicolas Humbert Emmanuel Boutant Pascal Didier Yves Mély Eleonore Real |
| author_sort | Nedal Taha |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-03-09T03:41:43Z |
| format | Article |
| id | doaj.art-b06fa11337784ce39d836b4094b87866 |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-03-09T03:41:43Z |
| publishDate | 2022-08-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Viruses |
| spelling | doaj.art-b06fa11337784ce39d836b4094b878662023-12-03T14:39:39ZengMDPI AGViruses1999-49152022-08-01148179810.3390/v14081798Upstream of N-Ras (Unr/CSDE1) Interacts with NCp7 and Gag, Modulating HIV-1 IRES-Mediated Translation InitiationNedal Taha0Sarwat Zgheib1Kamal Kant Sharma2Nicolas Humbert3Emmanuel Boutant4Pascal Didier5Yves Mély6Eleonore Real7UMR 7021 CNRS, Laboratoire de Bioimagerie et Pathologies, Université de Strasbourg, Faculté de Pharmacie, 67401 Illkirch-Graffenstaden, FranceUMR 7021 CNRS, Laboratoire de Bioimagerie et Pathologies, Université de Strasbourg, Faculté de Pharmacie, 67401 Illkirch-Graffenstaden, FranceUMR 7021 CNRS, Laboratoire de Bioimagerie et Pathologies, Université de Strasbourg, Faculté de Pharmacie, 67401 Illkirch-Graffenstaden, FranceUMR 7021 CNRS, Laboratoire de Bioimagerie et Pathologies, Université de Strasbourg, Faculté de Pharmacie, 67401 Illkirch-Graffenstaden, FranceUMR 7021 CNRS, Laboratoire de Bioimagerie et Pathologies, Université de Strasbourg, Faculté de Pharmacie, 67401 Illkirch-Graffenstaden, FranceUMR 7021 CNRS, Laboratoire de Bioimagerie et Pathologies, Université de Strasbourg, Faculté de Pharmacie, 67401 Illkirch-Graffenstaden, FranceUMR 7021 CNRS, Laboratoire de Bioimagerie et Pathologies, Université de Strasbourg, Faculté de Pharmacie, 67401 Illkirch-Graffenstaden, FranceUMR 7021 CNRS, Laboratoire de Bioimagerie et Pathologies, Université de Strasbourg, Faculté de Pharmacie, 67401 Illkirch-Graffenstaden, FranceThe 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.https://www.mdpi.com/1999-4915/14/8/1798HIVnucleocapsidNCp7UnrGagIRES |
| spellingShingle | Nedal Taha Sarwat Zgheib Kamal Kant Sharma Nicolas Humbert Emmanuel Boutant Pascal Didier Yves Mély Eleonore Real Upstream of N-Ras (Unr/CSDE1) Interacts with NCp7 and Gag, Modulating HIV-1 IRES-Mediated Translation Initiation Viruses HIV nucleocapsid NCp7 Unr Gag IRES |
| 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 | HIV nucleocapsid NCp7 Unr Gag IRES |
| url | https://www.mdpi.com/1999-4915/14/8/1798 |
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