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-Structure paper
タイトル | In situ structures of polymerase complex of mammalian reovirus illuminate RdRp activation and transcription regulation. |
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ジャーナル・号・ページ | Proc Natl Acad Sci U S A, Vol. 119, Issue 50, Page e2203054119, Year 2022 |
掲載日 | 2022年12月13日 |
著者 | Keyan Bao / Xueli Zhang / Dongyu Li / Wei Sun / Zhenzhao Sun / Jingfei Wang / Ping Zhu / |
PubMed 要旨 | Mammalian reovirus (reovirus) is a multilayered, turreted member of characterized by transcription of dsRNA genome within the innermost capsid shell. Here, we present high-resolution in situ ...Mammalian reovirus (reovirus) is a multilayered, turreted member of characterized by transcription of dsRNA genome within the innermost capsid shell. Here, we present high-resolution in situ structures of reovirus transcriptase complex in an intact double-layered virion, and in the uncoated single-layered core particles in the unloaded, reloaded, pre-elongation, and elongation states, respectively, obtained by cryo-electron microscopy and sub-particle reconstructions. At the template entry of RNA-dependent RNA polymerase (RdRp), the RNA-loading region gets flexible after uncoating resulting in the unloading of terminal genomic RNA and inactivity of transcription. However, upon adding transcriptional substrates, the RNA-loading region is recovered leading the RNAs loaded again. The priming loop in RdRp was found to play a critical role in regulating transcription, which hinders the elongation of transcript in virion and triggers the rearrangement of RdRp C-terminal domain (CTD) during elongation, resulting in splitting of template-transcript hybrid and opening of transcript exit. With the integration of these structures, a transcriptional model of reovirus with five states is proposed. Our structures illuminate the RdRp activation and regulation of the multilayered turreted reovirus. |
リンク | Proc Natl Acad Sci U S A / PubMed:36469786 / PubMed Central |
手法 | EM (単粒子) |
解像度 | 3.0 - 3.6 Å |
構造データ | EMDB-33770, PDB-7yed: EMDB-33778, PDB-7yev: EMDB-33779, PDB-7yez: EMDB-33780, PDB-7yf0: EMDB-33787, PDB-7yfe: |
化合物 | ChemComp-ZN: ChemComp-SAM: ChemComp-GTP: ChemComp-UTP: ChemComp-MG: |
由来 |
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キーワード | VIRAL PROTEIN/RNA / mammalian reovirus / cryo-em / RNA dependent RNA polymerase / transcription / VIRAL PROTEIN / VIRAL PROTEIN-RNA complex |