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Mutagenic Analysis of the HIV Restriction Factor Shiftless

The interferon-induced host cell protein shiftless (SFL) was reported to inhibit human immunodeficiency virus (HIV) infection by blocking the –1 programmed ribosomal frameshifting (–1PRF) required for expression of the Gag-Pol polyprotein. However, it is not clear how SFL inhibits –1PRF. To address...

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Autores principales: Jäger, Niklas, Ayyub, Shreya Ahana, Korniy, Natalia, Peske, Frank, Hoffmann, Markus, Rodnina, Marina V., Pöhlmann, Stefan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9324250/
https://www.ncbi.nlm.nih.gov/pubmed/35891432
http://dx.doi.org/10.3390/v14071454
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author Jäger, Niklas
Ayyub, Shreya Ahana
Korniy, Natalia
Peske, Frank
Hoffmann, Markus
Rodnina, Marina V.
Pöhlmann, Stefan
author_facet Jäger, Niklas
Ayyub, Shreya Ahana
Korniy, Natalia
Peske, Frank
Hoffmann, Markus
Rodnina, Marina V.
Pöhlmann, Stefan
author_sort Jäger, Niklas
collection PubMed
description The interferon-induced host cell protein shiftless (SFL) was reported to inhibit human immunodeficiency virus (HIV) infection by blocking the –1 programmed ribosomal frameshifting (–1PRF) required for expression of the Gag-Pol polyprotein. However, it is not clear how SFL inhibits –1PRF. To address this question, we focused on a 36 amino acids comprising region (termed required for antiviral activity (RAA)) that is essential for suppression of –1PRF and HIV infection and is missing from SFL short (SFLS), a splice variant of SFL with unknown function. Here, we confirm that SFL, but not SFLS, inhibits HIV –1PRF and show that inhibition is cell-type-independent. Mutagenic and biochemical analyses demonstrated that the RAA region is required for SFL self-interactions and confirmed that it is necessary for ribosome association and binding to the HIV RNA. Analysis of SFL mutants with six consecutive amino-acids-comprising deletions in the RAA region suggests effects on binding to the HIV RNA, complete inhibition of –1PRF, inhibition of Gag-Pol expression, and antiviral activity. In contrast, these amino acids did not affect SFL expression and were partially dispensable for SFL self-interactions and binding to the ribosome. Collectively, our results support the notion that SFL binds to the ribosome and the HIV RNA in order to block –1PRF and HIV infection, and suggest that the multimerization of SFL may be functionally important.
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spelling pubmed-93242502022-07-27 Mutagenic Analysis of the HIV Restriction Factor Shiftless Jäger, Niklas Ayyub, Shreya Ahana Korniy, Natalia Peske, Frank Hoffmann, Markus Rodnina, Marina V. Pöhlmann, Stefan Viruses Article The interferon-induced host cell protein shiftless (SFL) was reported to inhibit human immunodeficiency virus (HIV) infection by blocking the –1 programmed ribosomal frameshifting (–1PRF) required for expression of the Gag-Pol polyprotein. However, it is not clear how SFL inhibits –1PRF. To address this question, we focused on a 36 amino acids comprising region (termed required for antiviral activity (RAA)) that is essential for suppression of –1PRF and HIV infection and is missing from SFL short (SFLS), a splice variant of SFL with unknown function. Here, we confirm that SFL, but not SFLS, inhibits HIV –1PRF and show that inhibition is cell-type-independent. Mutagenic and biochemical analyses demonstrated that the RAA region is required for SFL self-interactions and confirmed that it is necessary for ribosome association and binding to the HIV RNA. Analysis of SFL mutants with six consecutive amino-acids-comprising deletions in the RAA region suggests effects on binding to the HIV RNA, complete inhibition of –1PRF, inhibition of Gag-Pol expression, and antiviral activity. In contrast, these amino acids did not affect SFL expression and were partially dispensable for SFL self-interactions and binding to the ribosome. Collectively, our results support the notion that SFL binds to the ribosome and the HIV RNA in order to block –1PRF and HIV infection, and suggest that the multimerization of SFL may be functionally important. MDPI 2022-06-30 /pmc/articles/PMC9324250/ /pubmed/35891432 http://dx.doi.org/10.3390/v14071454 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
Jäger, Niklas
Ayyub, Shreya Ahana
Korniy, Natalia
Peske, Frank
Hoffmann, Markus
Rodnina, Marina V.
Pöhlmann, Stefan
Mutagenic Analysis of the HIV Restriction Factor Shiftless
title Mutagenic Analysis of the HIV Restriction Factor Shiftless
title_full Mutagenic Analysis of the HIV Restriction Factor Shiftless
title_fullStr Mutagenic Analysis of the HIV Restriction Factor Shiftless
title_full_unstemmed Mutagenic Analysis of the HIV Restriction Factor Shiftless
title_short Mutagenic Analysis of the HIV Restriction Factor Shiftless
title_sort mutagenic analysis of the hiv restriction factor shiftless
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9324250/
https://www.ncbi.nlm.nih.gov/pubmed/35891432
http://dx.doi.org/10.3390/v14071454
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