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Title | Structural insights into viral genome replication by the severe fever with thrombocytopenia syndrome virus L protein. |
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Journal, issue, pages | Nucleic Acids Res, Vol. 51, Issue 3, Page 1424-1442, Year 2023 |
Publish date | Feb 22, 2023 |
Authors | Harry M Williams / Sigurdur R Thorkelsson / Dominik Vogel / Morlin Milewski / Carola Busch / Stephen Cusack / Kay Grünewald / Emmanuelle R J Quemin / Maria Rosenthal / |
PubMed Abstract | Severe fever with thrombocytopenia syndrome virus (SFTSV) is a phenuivirus that has rapidly become endemic in several East Asian countries. The large (L) protein of SFTSV, which includes the RNA- ...Severe fever with thrombocytopenia syndrome virus (SFTSV) is a phenuivirus that has rapidly become endemic in several East Asian countries. The large (L) protein of SFTSV, which includes the RNA-dependent RNA polymerase (RdRp), is responsible for catalysing viral genome replication and transcription. Here, we present 5 cryo-electron microscopy (cryo-EM) structures of the L protein in several states of the genome replication process, from pre-initiation to late-stage elongation, at a resolution of up to 2.6 Å. We identify how the L protein binds the 5' viral RNA in a hook-like conformation and show how the distal 5' and 3' RNA ends form a duplex positioning the 3' RNA terminus in the RdRp active site ready for initiation. We also observe the L protein stalled in the early and late stages of elongation with the RdRp core accommodating a 10-bp product-template duplex. This duplex ultimately splits with the template binding to a designated 3' secondary binding site. The structural data and observations are complemented by in vitro biochemical and cell-based mini-replicon assays. Altogether, our data provide novel key insights into the mechanism of viral genome replication by the SFTSV L protein and will aid drug development against segmented negative-strand RNA viruses. |
External links | Nucleic Acids Res / PubMed:36651274 / PubMed Central |
Methods | EM (single particle) |
Resolution | 2.6 - 3.6 Å |
Structure data | EMDB-15607, PDB-8as6: EMDB-15608, PDB-8as7: EMDB-15610, PDB-8asb: EMDB-15614, PDB-8asd: EMDB-15615, PDB-8asg: |
Chemicals | ChemComp-MG: ChemComp-EPE: ChemComp-2KH: ChemComp-HOH: |
Source |
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Keywords | VIRAL PROTEIN / SFTSV RNA-DEPENDENT RNA POLYMERASE / VIRAL RNA |