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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MDPI AG
2022-06-01
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| Series: | Viruses |
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| Online Access: | https://www.mdpi.com/1999-4915/14/7/1454 |
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| author | Niklas Jäger Shreya Ahana Ayyub Natalia Korniy Frank Peske Markus Hoffmann Marina V. Rodnina Stefan Pöhlmann |
| author_facet | Niklas Jäger Shreya Ahana Ayyub Natalia Korniy Frank Peske Markus Hoffmann Marina V. Rodnina Stefan Pöhlmann |
| author_sort | Niklas Jäger |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-03-09T05:43:34Z |
| format | Article |
| id | doaj.art-d1707e53f5944476bd73b808472c87ef |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-03-09T05:43:34Z |
| publishDate | 2022-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Viruses |
| spelling | doaj.art-d1707e53f5944476bd73b808472c87ef2023-12-03T12:23:10ZengMDPI AGViruses1999-49152022-06-01147145410.3390/v14071454Mutagenic Analysis of the HIV Restriction Factor ShiftlessNiklas Jäger0Shreya Ahana Ayyub1Natalia Korniy2Frank Peske3Markus Hoffmann4Marina V. Rodnina5Stefan Pöhlmann6Infection Biology Unit, German Primate Center, 37077 Göttingen, GermanyMax Planck Institute for Multidisciplinary Sciences, 37077 Göttingen, GermanyMax Planck Institute for Multidisciplinary Sciences, 37077 Göttingen, GermanyMax Planck Institute for Multidisciplinary Sciences, 37077 Göttingen, GermanyInfection Biology Unit, German Primate Center, 37077 Göttingen, GermanyMax Planck Institute for Multidisciplinary Sciences, 37077 Göttingen, GermanyInfection Biology Unit, German Primate Center, 37077 Göttingen, GermanyThe 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.https://www.mdpi.com/1999-4915/14/7/1454–1 programmed ribosomal frameshiftingshiftlessC19orf66restriction factorHIV-1 |
| spellingShingle | Niklas Jäger Shreya Ahana Ayyub Natalia Korniy Frank Peske Markus Hoffmann Marina V. Rodnina Stefan Pöhlmann Mutagenic Analysis of the HIV Restriction Factor Shiftless Viruses –1 programmed ribosomal frameshifting shiftless C19orf66 restriction factor HIV-1 |
| 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 | –1 programmed ribosomal frameshifting shiftless C19orf66 restriction factor HIV-1 |
| url | https://www.mdpi.com/1999-4915/14/7/1454 |
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