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Mol Cell. 2015 Mar 5;57(5):925-935. doi: 10.1016/j.molcel.2014.12.031. Epub 2015 Jan 22.

Cryo-EM structure of influenza virus RNA polymerase complex at 4.3 Å resolution.

Author information

1
National Key Laboratory of Biomacromolecules, University of Chinese Academy of Sciences, Institute of Biophysics, Chinese Academy of Sciences,Beijing, China 100101.
2
Ministry of Education Key Laboratory of Protein Science, Tsinghua-Peking Joint Center for Life Sciences, Center for Structural Biology, School of Life Sciences, Tsinghua University, Beijing, China, 100084.
3
Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, USA CA90095; Center of System Medicine, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, China 100005.
4
Swiss Light Source, Paul Scherrer Institute, Villigen, Switzerland CH-5232.
5
The State Key Laboratory for DTID, First Affiliated Hospital, College of Medicine, Zhejiang University; Collaborative Innovation Center for DTID, 79 Qingchun Road, Hangzhou 310003, China.
6
Ministry of Education Key Laboratory of Protein Science, Tsinghua-Peking Joint Center for Life Sciences, Center for Structural Biology, School of Life Sciences, Tsinghua University, Beijing, China, 100084. Electronic address: hongweiwang@tsinghua.edu.cn.
7
National Key Laboratory of Biomacromolecules, University of Chinese Academy of Sciences, Institute of Biophysics, Chinese Academy of Sciences,Beijing, China 100101; The State Key Laboratory for DTID, First Affiliated Hospital, College of Medicine, Zhejiang University; Collaborative Innovation Center for DTID, 79 Qingchun Road, Hangzhou 310003, China. Electronic address: liuy@ibp.ac.cn.

Abstract

Replication and transcription of influenza virus genome mainly depend on its RNA-dependent RNA polymerase (RdRP), composed of the PA, PB1, and PB2 subunits. Although extensively studied, the underlying mechanism of the RdRP complex is still unclear. Here we report the biochemical characterization of influenza RdRP subcomplex comprising PA, PB1, and N terminus of PB2, which exist as dimer in solution and can assemble into a tetramer state, regulated by vRNA promoter. Using single-particle cryo-electron microscopy, we have reconstructed the RdRP tetramer complex at 4.3 Å, highlighting the assembly and interfaces between monomers within the tetrameric structure. The individual RdRP subcomplex contains all the characterized motifs and appears as a cage-like structure. High-throughput mutagenesis profiling revealed that residues involved in the oligomer state formation are critical for viral life cycle. Our results lay a solid base for understanding the mechanism of replication of influenza and other negative-stranded RNA viruses.

PMID:
25620561
DOI:
10.1016/j.molcel.2014.12.031
[Indexed for MEDLINE]
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