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Structure of RdRps within a transcribing dsRNA virus provides insights into the mechanisms of RNA synthesis.
J Mol Biol 2019JM

Abstract

RNA-dependent RNA polymerases (RdRps) catalyze RNA synthesis of RNA viruses. During initiation of RNA synthesis, the RdRp catalyzes the formation of the first dinucleotide, acting as primer for subsequent processive RNA elongation. Here, we present the structure of the RdRp complexes in the dinucleotide primed state in situ within a transcribing cypovirus under near physiological conditions using cryo-electron microscopy. The 3' end of RNA templates, paired RNA dinucleotide primer, incoming nucleotide, and catalytic divalent cations in the RdRp were resolved at 3.8 Å resolution. The end of the RNA template and the dinucleotide is buttressed by the aromatic tyrosine in a loop from the RdRp bracelet domain. Our structure reveals the interactions between the nucleotide substrates and the conserved residues during the RdRp initiation, and the coordinated structural changes preceding the elongation stage. In addition, it provides the direct evidence for existence of the slow step of the dinucleotide primed state in the viral RdRp transcription.

Authors+Show Affiliations

College of Physics and Information Science, Key Laboratory of Low-dimensional Quantum Structures, And Quantum Control of the Ministry of Education, Synergetic Innovation Center for Quantum Effects and Applications, Hunan Normal University, Changsha, China; Technology Center for Protein Sciences, School of Life Sciences, Tsinghua University, Beijing, China.State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China.College of Physics and Information Science, Key Laboratory of Low-dimensional Quantum Structures, And Quantum Control of the Ministry of Education, Synergetic Innovation Center for Quantum Effects and Applications, Hunan Normal University, Changsha, China.College of Physics and Information Science, Key Laboratory of Low-dimensional Quantum Structures, And Quantum Control of the Ministry of Education, Synergetic Innovation Center for Quantum Effects and Applications, Hunan Normal University, Changsha, China.Technology Center for Protein Sciences, School of Life Sciences, Tsinghua University, Beijing, China.State Key Laboratory of Infectious Disease Prevention and Control, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, China. Electronic address: songjd@ivdc.chinacdc.cn.College of Physics and Information Science, Key Laboratory of Low-dimensional Quantum Structures, And Quantum Control of the Ministry of Education, Synergetic Innovation Center for Quantum Effects and Applications, Hunan Normal University, Changsha, China. Electronic address: hrliu@hunnu.edu.cn.Technology Center for Protein Sciences, School of Life Sciences, Tsinghua University, Beijing, China. Electronic address: lingpengcheng@mail.tsinghua.edu.cn.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

31629769

Citation

Li, Xiaowu, et al. "Structure of RdRps Within a Transcribing dsRNA Virus Provides Insights Into the Mechanisms of RNA Synthesis." Journal of Molecular Biology, 2019.
Li X, Wang L, Wang X, et al. Structure of RdRps within a transcribing dsRNA virus provides insights into the mechanisms of RNA synthesis. J Mol Biol. 2019.
Li, X., Wang, L., Wang, X., Chen, W., Yang, T., Song, J., ... Cheng, L. (2019). Structure of RdRps within a transcribing dsRNA virus provides insights into the mechanisms of RNA synthesis. Journal of Molecular Biology, doi:10.1016/j.jmb.2019.09.015.
Li X, et al. Structure of RdRps Within a Transcribing dsRNA Virus Provides Insights Into the Mechanisms of RNA Synthesis. J Mol Biol. 2019 Oct 17; PubMed PMID: 31629769.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Structure of RdRps within a transcribing dsRNA virus provides insights into the mechanisms of RNA synthesis. AU - Li,Xiaowu, AU - Wang,Li, AU - Wang,Xurong, AU - Chen,Wenyuan, AU - Yang,Tao, AU - Song,Jingdong, AU - Liu,Hongrong, AU - Cheng,Lingpeng, Y1 - 2019/10/17/ PY - 2019/08/05/received PY - 2019/09/11/revised PY - 2019/09/16/accepted PY - 2019/10/21/entrez PY - 2019/10/21/pubmed PY - 2019/10/21/medline KW - RNA synthesis KW - RNA virus KW - RNA-dependent RNA polymerase KW - cryo-EM KW - transcription JF - Journal of molecular biology JO - J. Mol. Biol. N2 - RNA-dependent RNA polymerases (RdRps) catalyze RNA synthesis of RNA viruses. During initiation of RNA synthesis, the RdRp catalyzes the formation of the first dinucleotide, acting as primer for subsequent processive RNA elongation. Here, we present the structure of the RdRp complexes in the dinucleotide primed state in situ within a transcribing cypovirus under near physiological conditions using cryo-electron microscopy. The 3' end of RNA templates, paired RNA dinucleotide primer, incoming nucleotide, and catalytic divalent cations in the RdRp were resolved at 3.8 Å resolution. The end of the RNA template and the dinucleotide is buttressed by the aromatic tyrosine in a loop from the RdRp bracelet domain. Our structure reveals the interactions between the nucleotide substrates and the conserved residues during the RdRp initiation, and the coordinated structural changes preceding the elongation stage. In addition, it provides the direct evidence for existence of the slow step of the dinucleotide primed state in the viral RdRp transcription. SN - 1089-8638 UR - https://www.unboundmedicine.com/medline/citation/31629769/Structure_of_RdRps_within_a_transcribing_dsRNA_virus_provides_insights_into_the_mechanisms_of_RNA_synthesis L2 - https://linkinghub.elsevier.com/retrieve/pii/S0022-2836(19)30570-4 DB - PRIME DP - Unbound Medicine ER -