The role of the priming loop in influenza A virus RNA synthesis
RNA-dependent RNA polymerases (RdRps) are used by RNA viruses to replicate and transcribe their RNA genomes1 . They adopt a closed, right-handed fold with conserved subdomains called palm, fingers and thumb1,2. Conserved RdRp motifs A–F coordinate the viral RNA template, NTPs and magnesium ions to f...
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| Fformat: | Journal article |
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Nature
2016
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| _version_ | 1826286617926565888 |
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| author | Te Velthuis, A Robb, N Kapanidis, A Fodor, E |
| author_facet | Te Velthuis, A Robb, N Kapanidis, A Fodor, E |
| author_sort | Te Velthuis, A |
| collection | OXFORD |
| description | RNA-dependent RNA polymerases (RdRps) are used by RNA viruses to replicate and transcribe their RNA genomes1 . They adopt a closed, right-handed fold with conserved subdomains called palm, fingers and thumb1,2. Conserved RdRp motifs A–F coordinate the viral RNA template, NTPs and magnesium ions to facilitate nucleotide condensation1 . For the initiation of RNA synthesis, most RdRps use either a primer-dependent or de novo mechanism3. The influenza A virus RdRp, in contrast, uses a capped RNA oligonucleotide to initiate transcription, and a combination of terminal and internal de novo initiation for replication4. To understand how the influenza A virus RdRp coordinates these processes, we analysed the function of a thumb subdomain β-hairpin using initiation, elongation and single-molecule Förster resonance energy transfer (sm-FRET) assays. Our data indicate that this β-hairpin is essential for terminal initiation during replication, but not necessary for internal initiation and transcription. Analysis of individual residues in the tip of the β-hairpin shows that PB1 proline 651 is critical for efficient RNA synthesis in vitro and in cell culture. Overall, this work advances our understanding of influenza A virus RNA synthesis and identifies the initiation platform of viral replication. |
| first_indexed | 2024-03-07T01:46:24Z |
| format | Journal article |
| id | oxford-uuid:989253eb-b80c-4a63-92bb-72dcad67bc8f |
| institution | University of Oxford |
| last_indexed | 2024-03-07T01:46:24Z |
| publishDate | 2016 |
| publisher | Nature |
| record_format | dspace |
| spelling | oxford-uuid:989253eb-b80c-4a63-92bb-72dcad67bc8f2022-03-27T00:07:57ZThe role of the priming loop in influenza A virus RNA synthesisJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:989253eb-b80c-4a63-92bb-72dcad67bc8fSymplectic Elements at OxfordNature2016Te Velthuis, ARobb, NKapanidis, AFodor, ERNA-dependent RNA polymerases (RdRps) are used by RNA viruses to replicate and transcribe their RNA genomes1 . They adopt a closed, right-handed fold with conserved subdomains called palm, fingers and thumb1,2. Conserved RdRp motifs A–F coordinate the viral RNA template, NTPs and magnesium ions to facilitate nucleotide condensation1 . For the initiation of RNA synthesis, most RdRps use either a primer-dependent or de novo mechanism3. The influenza A virus RdRp, in contrast, uses a capped RNA oligonucleotide to initiate transcription, and a combination of terminal and internal de novo initiation for replication4. To understand how the influenza A virus RdRp coordinates these processes, we analysed the function of a thumb subdomain β-hairpin using initiation, elongation and single-molecule Förster resonance energy transfer (sm-FRET) assays. Our data indicate that this β-hairpin is essential for terminal initiation during replication, but not necessary for internal initiation and transcription. Analysis of individual residues in the tip of the β-hairpin shows that PB1 proline 651 is critical for efficient RNA synthesis in vitro and in cell culture. Overall, this work advances our understanding of influenza A virus RNA synthesis and identifies the initiation platform of viral replication. |
| spellingShingle | Te Velthuis, A Robb, N Kapanidis, A Fodor, E The role of the priming loop in influenza A virus RNA synthesis |
| title | The role of the priming loop in influenza A virus RNA synthesis |
| title_full | The role of the priming loop in influenza A virus RNA synthesis |
| title_fullStr | The role of the priming loop in influenza A virus RNA synthesis |
| title_full_unstemmed | The role of the priming loop in influenza A virus RNA synthesis |
| title_short | The role of the priming loop in influenza A virus RNA synthesis |
| title_sort | role of the priming loop in influenza a virus rna synthesis |
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