title: Cryo-EM structure of influenza virus RNA polymerase complex at 4.3 Å resolution.
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.
authors.affiliation: National Key Laboratory of Biomacromolecules, University of Chinese Academy of Sciences, Institute of Biophysics, Chinese Academy of Sciences,Beijing, China 100101.
authors.forename: Shenghai
authors.initials: S
authors.lastname: Chang
authors.affiliation: National Key Laboratory of Biomacromolecules, University of Chinese Academy of Sciences, Institute of Biophysics, Chinese Academy of Sciences,Beijing, China 100101.
authors.forename: Dapeng
authors.initials: D
authors.lastname: Sun
authors.affiliation: National Key Laboratory of Biomacromolecules, University of Chinese Academy of Sciences, Institute of Biophysics, Chinese Academy of Sciences,Beijing, China 100101.
authors.forename: Huanhuan
authors.initials: H
authors.lastname: Liang
authors.affiliation: 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.
authors.forename: Jia
authors.initials: J
authors.lastname: Wang
authors.affiliation: National Key Laboratory of Biomacromolecules, University of Chinese Academy of Sciences, Institute of Biophysics, Chinese Academy of Sciences,Beijing, China 100101.
authors.forename: Jun
authors.initials: J
authors.lastname: Li
authors.affiliation: National Key Laboratory of Biomacromolecules, University of Chinese Academy of Sciences, Institute of Biophysics, Chinese Academy of Sciences,Beijing, China 100101.
authors.forename: Lu
authors.initials: L
authors.lastname: Guo
authors.affiliation: National Key Laboratory of Biomacromolecules, University of Chinese Academy of Sciences, Institute of Biophysics, Chinese Academy of Sciences,Beijing, China 100101.
authors.forename: Xiangli
authors.initials: X
authors.lastname: Wang
authors.affiliation: National Key Laboratory of Biomacromolecules, University of Chinese Academy of Sciences, Institute of Biophysics, Chinese Academy of Sciences,Beijing, China 100101.
authors.forename: Chengcheng
authors.initials: C
authors.lastname: Guan
authors.affiliation: National Key Laboratory of Biomacromolecules, University of Chinese Academy of Sciences, Institute of Biophysics, Chinese Academy of Sciences,Beijing, China 100101.
authors.forename: Bhargavi M
authors.initials: BM
authors.lastname: Boruah
authors.affiliation: National Key Laboratory of Biomacromolecules, University of Chinese Academy of Sciences, Institute of Biophysics, Chinese Academy of Sciences,Beijing, China 100101.
authors.forename: Lingmin
authors.initials: L
authors.lastname: Yuan
authors.affiliation: National Key Laboratory of Biomacromolecules, University of Chinese Academy of Sciences, Institute of Biophysics, Chinese Academy of Sciences,Beijing, China 100101.
authors.forename: Feng
authors.initials: F
authors.lastname: Feng
authors.affiliation: National Key Laboratory of Biomacromolecules, University of Chinese Academy of Sciences, Institute of Biophysics, Chinese Academy of Sciences,Beijing, China 100101.
authors.forename: Mingrui
authors.initials: M
authors.lastname: Yang
authors.affiliation: 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.
authors.forename: Lulan
authors.initials: L
authors.lastname: Wang
authors.affiliation: 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.
authors.forename: Yao
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Swiss Light Source, Paul Scherrer Institute, Villigen, Switzerland CH-5232.
authors.forename: Justyna
authors.initials: J
authors.lastname: Wojdyla
authors.affiliation: 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.
authors.forename: Lanjuan
authors.initials: L
authors.lastname: Li
authors.affiliation: 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.
authors.forename: Jiawei
authors.initials: J
authors.lastname: Wang
authors.affiliation: Swiss Light Source, Paul Scherrer Institute, Villigen, Switzerland CH-5232.
authors.forename: Meitian
authors.initials: M
authors.lastname: Wang
authors.affiliation: 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.
authors.forename: Genhong
authors.initials: G
authors.lastname: Cheng
authors.affiliation: 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.
authors.forename: Hong-Wei
authors.initials: HW
authors.lastname: Wang
authors.affiliation: 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.
authors.forename: Yingfang
authors.initials: Y
authors.lastname: Liu
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D020285:Cryoelectron Microscopy / Q000379:methods*; D057809:HEK293 Cells; D006801:Humans; D021621:Imaging, Three-Dimensional; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation; D009975:Orthomyxoviridae / Q000201:enzymology* / Q000235:genetics; D055503:Protein Multimerization; D020836:Protein Structure, Quaternary; D017434:Protein Structure, Tertiary; D021122:Protein Subunits / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012324:RNA-Dependent RNA Polymerase / Q000737:chemistry / Q000235:genetics / Q000648:ultrastructure*; D017386:Sequence Homology, Amino Acid; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics / Q000648:ultrastructure*
chemical_list: D021122:Protein Subunits; D014764:Viral Proteins; D012324:RNA-Dependent RNA Polymerase