German Federal Ministry for Education and Research
01KI2019
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ジャーナル: Nucleic Acids Res / 年: 2023 タイトル: Structural insights into viral genome replication by the severe fever with thrombocytopenia syndrome virus L protein. 著者: Harry M Williams / Sigurdur R Thorkelsson / Dominik Vogel / Morlin Milewski / Carola Busch / Stephen Cusack / Kay Grünewald / Emmanuelle R J Quemin / Maria Rosenthal / 要旨: 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.
名称: RNA (5'-R(P*AP*AP*AP*AP*AP*AP*GP*AP*UP*CP*UP*GP*GP*GP*CP*GP*GP*UP*CP*UP*UP*UP*GP*UP*GP*U)-3') タイプ: rna / ID: 2 詳細: Additional 6A's added to template RNA (nt 21 - 26). Start of 3' cRNA bound to secondary binding site, remainder defined in the L protein core - nts in between are not clearly defined in the ...詳細: Additional 6A's added to template RNA (nt 21 - 26). Start of 3' cRNA bound to secondary binding site, remainder defined in the L protein core - nts in between are not clearly defined in the EM map (although clearly there in the blurred map) and so not fit in the PDB. コピー数: 1
名称: RNA (5'-R(P*AP*CP*AP*CP*AP*AP*AP*GP*AP*CP*CP*GP*CP*CP*CP*AP*GP*AP*CP*CP*UP*UP*UP*UP*UP*U)-3') タイプ: rna / ID: 3 詳細: Additional 6A's added to template RNA (nt 21 - 26) so poly-U stretch here is artificial. At position 19 on the 3' cRNA we have a A. However, our investigations show that a C is mis- ...詳細: Additional 6A's added to template RNA (nt 21 - 26) so poly-U stretch here is artificial. At position 19 on the 3' cRNA we have a A. However, our investigations show that a C is mis-incorporated at this point in the growing strand. If I place the correct growing strand sequence in the box, it is flagged as incorrect because in our structure we have C instead of the expected A. To get around this, I have changed the sample sequence to reflect what we see in our experimental setup. コピー数: 1