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Single-molecule FRET reveals the pre-initiation and initiation conformations of influenza virus promoter RNA
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Robb, Nicole C., te Velthuis, Aartjan J. W., Wieneke, Ralph, Tampé, Robert, Cordes, Thorben, Fodor, Ervin and Kapanidis, Achillefs N. (2016) Single-molecule FRET reveals the pre-initiation and initiation conformations of influenza virus promoter RNA. Nucleic Acids Research, 44 (21). pp. 10304-10315. doi:10.1093/nar/gkw884
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Official URL: http://dx.doi.org/10.1093/nar/gkw884
Abstract
Influenza viruses have a segmented viral RNA (vRNA) genome, which is replicated by the viral RNA-dependent RNA polymerase (RNAP). Replication initiates on the vRNA 3΄ terminus, producing a complementary RNA (cRNA) intermediate, which serves as a template for the synthesis of new vRNA. RNAP structures show the 3΄ terminus of the vRNA template in a pre-initiation state, bound on the surface of the RNAP rather than in the active site; no information is available on 3΄ cRNA binding. Here, we have used single-molecule Förster resonance energy transfer (smFRET) to probe the viral RNA conformations that occur during RNAP binding and initial replication. We show that even in the absence of nucleotides, the RNAP-bound 3΄ termini of both vRNA and cRNA exist in two conformations, corresponding to the pre-initiation state and an initiation conformation in which the 3΄ terminus of the viral RNA is in the RNAP active site. Nucleotide addition stabilises the 3΄ vRNA in the active site and results in unwinding of the duplexed region of the promoter. Our data provide insights into the dynamic motions of RNA that occur during initial influenza replication and has implications for our understanding of the replication mechanisms of similar pathogenic viruses.
| Item Type: | Journal Article | |||||||||||||||||||||||||||||||||||||||
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| Subjects: | Q Science > QC Physics Q Science > QP Physiology Q Science > QR Microbiology |
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| Divisions: | Faculty of Medicine > Warwick Medical School > Biomedical Sciences > Cell & Developmental Biology Faculty of Medicine > Warwick Medical School > Biomedical Sciences Faculty of Medicine > Warwick Medical School |
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| Library of Congress Subject Headings (LCSH): | Influenza viruses, Influenza viruses -- Reproduction , RNA, Energy transfer, Molecular dynamics | |||||||||||||||||||||||||||||||||||||||
| Journal or Publication Title: | Nucleic Acids Research | |||||||||||||||||||||||||||||||||||||||
| Publisher: | Oxford University Press | |||||||||||||||||||||||||||||||||||||||
| ISSN: | 0305-1048 | |||||||||||||||||||||||||||||||||||||||
| Official Date: | December 2016 | |||||||||||||||||||||||||||||||||||||||
| Dates: |
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| Date of first compliant deposit: | 28 July 2020 | |||||||||||||||||||||||||||||||||||||||
| Volume: | 44 | |||||||||||||||||||||||||||||||||||||||
| Number: | 21 | |||||||||||||||||||||||||||||||||||||||
| Page Range: | pp. 10304-10315 | |||||||||||||||||||||||||||||||||||||||
| DOI: | 10.1093/nar/gkw884 | |||||||||||||||||||||||||||||||||||||||
| Status: | Peer Reviewed | |||||||||||||||||||||||||||||||||||||||
| Publication Status: | Published | |||||||||||||||||||||||||||||||||||||||
| Access rights to Published version: | Open Access | |||||||||||||||||||||||||||||||||||||||
| RIOXX Funder/Project Grant: |
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