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Cited by 50Viral Infections and VectorsVirology and Viral DiseasesViral Infections and Outbreaks Research

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Conformational changes in Lassa virus L protein associated with promoter binding and RNA synthesis activity

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Kouba, Tomas, Vogel, Dominik, Thorkelsson, Sigurdur R., Quemin, Emmanuelle R. J., Williams, Harry M., Milewski, Morlin, Busch, Carola, Günther, Stephan, Grünewald, Kay, Rosenthal, Maria, Cusack, Stephen·2021·Nature Communications
DOI 10.1038/s41467-021-27305-5PMC PMC8639829

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Abstract

Lassa virus is endemic in West Africa and can cause severe hemorrhagic fever. The viral L protein transcribes and replicates the RNA genome via its RNA-dependent RNA polymerase activity. Here, we present nine cryo-EM structures of the L protein in the apo-, promoter-bound pre-initiation and active RNA synthesis states. We characterize distinct binding pockets for the conserved 3’ and 5’ promoter RNAs and show how full-promoter binding induces a distinct pre-initiation conformation. In the apo- and early elongation states, the endonuclease is inhibited by two distinct L protein peptides, whereas in the pre-initiation state it is uninhibited. In the early elongation state, a template-product duplex is bound in the active site cavity together with an incoming non-hydrolysable nucleotide and the full C-terminal region of the L protein, including the putative cap-binding domain, is well-ordered. These data advance our mechanistic understanding of how this flexible and multifunctional molecular machine is activated.

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Metadata

Authors
Kouba, T., Vogel, D., Thorkelsson, S. R., Quemin, E. R. J., Williams, H. M., Milewski, M., Busch, C., Günther, S., Grünewald, K., Rosenthal, M., & Cusack, S.
Affiliations
  • Bernhard Nocht Institute for Tropical Medicine (DE)
  • Leibniz Institute of Virology (LIV) (DE)
  • European Molecular Biology Laboratory (FR)
Corresponding author
  • Rosenthal, Maria
  • Cusack, Stephen
ORCID
Venue
Nature Communications
Publisher
Springer Science and Business Media LLC
Published
2021-12-02
Volume
12
Issue
1
Pages
7018-7018
ISSN
2041-1723 (ISSN-L)
DOI
10.1038/s41467-021-27305-5
PMC
PMC8639829
OpenAlex
W3216452123
Cited by
50
References
1
Citations by year
2026: 9 · 2025: 9 · 2024: 10 · 2023: 15 · 2022: 6 · 2021: 1
FWCI
2.53
Type
journal article
Language
en
Open access
Licence
Locations
  • Nature Communications (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
  • Leibniz-Gemeinschaft (K72/2017)
  • Bundesministerium für Bildung und Forschung (01KI2019)
  • Deutsche Forschungsgemeinschaft (INST 152/772-1 and 774-1)
  • Alexander von Humboldt-Stiftung
Topics
  • Health Sciences › Medicine › Infectious Diseases › Viral Infections and Vectors
  • Health Sciences › Medicine › Epidemiology › Virology and Viral Diseases
  • Health Sciences › Medicine › Infectious Diseases › Viral Infections and Outbreaks Research
Keywords
Lassa virus, L protein, RNA-dependent RNA polymerase, promoter binding, RNA synthesis, preinitiation complex, conformational change, endonuclease inhibition, viral RNA replication, hemorrhagic fever
MeSH terms
Protein Conformation, alpha-Helical, Protein Conformation, beta-Strand, Cloning, Molecular, Escherichia coli, Genetic Vectors, Lassa virus, Models, Molecular, Promoter Regions, Genetic, Protein Binding, Recombinant Proteins, RNA-Dependent RNA Polymerase, RNA, Viral, Substrate Specificity, Transcription, Genetic, Viral Proteins, Gene Expression, Catalytic Domain, Amino Acid Motifs, Protein Interaction Domains and Motifs

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