Modulation of Viral Programmed Ribosomal Frameshifting and Stop Codon Readthrough by the Host Restriction Factor Shiftless
The product of the interferon-stimulated gene <i>C19orf66</i>, Shiftless (SHFL)<i>,</i> restricts human immunodeficiency virus replication through downregulation of the efficiency of the viral <i>gag/pol</i> frameshifting signal. In this study, we demonstrate that...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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MDPI AG
2021-06-01
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| Series: | Viruses |
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| Online Access: | https://www.mdpi.com/1999-4915/13/7/1230 |
| _version_ | 1797411979084693504 |
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| author | Sawsan Napthine Chris H. Hill Holly C. M. Nugent Ian Brierley |
| author_facet | Sawsan Napthine Chris H. Hill Holly C. M. Nugent Ian Brierley |
| author_sort | Sawsan Napthine |
| collection | DOAJ |
| description | The product of the interferon-stimulated gene <i>C19orf66</i>, Shiftless (SHFL)<i>,</i> restricts human immunodeficiency virus replication through downregulation of the efficiency of the viral <i>gag/pol</i> frameshifting signal. In this study, we demonstrate that bacterially expressed, purified SHFL can decrease the efficiency of programmed ribosomal frameshifting in vitro at a variety of sites, including the RNA pseudoknot-dependent signals of the coronaviruses IBV, SARS-CoV and SARS-CoV-2, and the protein-dependent stimulators of the cardioviruses EMCV and TMEV. SHFL also reduced the efficiency of stop-codon readthrough at the murine leukemia virus <i>gag/pol</i> signal. Using size-exclusion chromatography, we confirm the binding of the purified protein to mammalian ribosomes in vitro. Finally, through electrophoretic mobility shift assays and mutational analysis, we show that expressed SHFL has strong RNA binding activity that is necessary for full activity in the inhibition of frameshifting, but shows no clear specificity for stimulatory RNA structures. |
| first_indexed | 2024-03-09T04:54:22Z |
| format | Article |
| id | doaj.art-af04608523794b58871706c821d41b33 |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-03-09T04:54:22Z |
| publishDate | 2021-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Viruses |
| spelling | doaj.art-af04608523794b58871706c821d41b332023-12-03T13:08:01ZengMDPI AGViruses1999-49152021-06-01137123010.3390/v13071230Modulation of Viral Programmed Ribosomal Frameshifting and Stop Codon Readthrough by the Host Restriction Factor ShiftlessSawsan Napthine0Chris H. Hill1Holly C. M. Nugent2Ian Brierley3Department of Pathology, University of Cambridge, Cambridge CB2 1QP, UKDepartment of Pathology, University of Cambridge, Cambridge CB2 1QP, UKDepartment of Pathology, University of Cambridge, Cambridge CB2 1QP, UKDepartment of Pathology, University of Cambridge, Cambridge CB2 1QP, UKThe product of the interferon-stimulated gene <i>C19orf66</i>, Shiftless (SHFL)<i>,</i> restricts human immunodeficiency virus replication through downregulation of the efficiency of the viral <i>gag/pol</i> frameshifting signal. In this study, we demonstrate that bacterially expressed, purified SHFL can decrease the efficiency of programmed ribosomal frameshifting in vitro at a variety of sites, including the RNA pseudoknot-dependent signals of the coronaviruses IBV, SARS-CoV and SARS-CoV-2, and the protein-dependent stimulators of the cardioviruses EMCV and TMEV. SHFL also reduced the efficiency of stop-codon readthrough at the murine leukemia virus <i>gag/pol</i> signal. Using size-exclusion chromatography, we confirm the binding of the purified protein to mammalian ribosomes in vitro. Finally, through electrophoretic mobility shift assays and mutational analysis, we show that expressed SHFL has strong RNA binding activity that is necessary for full activity in the inhibition of frameshifting, but shows no clear specificity for stimulatory RNA structures.https://www.mdpi.com/1999-4915/13/7/1230ribosomeframeshiftreadthroughShiftlessvirus |
| spellingShingle | Sawsan Napthine Chris H. Hill Holly C. M. Nugent Ian Brierley Modulation of Viral Programmed Ribosomal Frameshifting and Stop Codon Readthrough by the Host Restriction Factor Shiftless Viruses ribosome frameshift readthrough Shiftless virus |
| title | Modulation of Viral Programmed Ribosomal Frameshifting and Stop Codon Readthrough by the Host Restriction Factor Shiftless |
| title_full | Modulation of Viral Programmed Ribosomal Frameshifting and Stop Codon Readthrough by the Host Restriction Factor Shiftless |
| title_fullStr | Modulation of Viral Programmed Ribosomal Frameshifting and Stop Codon Readthrough by the Host Restriction Factor Shiftless |
| title_full_unstemmed | Modulation of Viral Programmed Ribosomal Frameshifting and Stop Codon Readthrough by the Host Restriction Factor Shiftless |
| title_short | Modulation of Viral Programmed Ribosomal Frameshifting and Stop Codon Readthrough by the Host Restriction Factor Shiftless |
| title_sort | modulation of viral programmed ribosomal frameshifting and stop codon readthrough by the host restriction factor shiftless |
| topic | ribosome frameshift readthrough Shiftless virus |
| url | https://www.mdpi.com/1999-4915/13/7/1230 |
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