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Structure paper

TitleStructural and molecular basis for Cardiovirus 2A protein as a viral gene expression switch.
Journal, issue, pagesNat Commun, Vol. 12, Issue 1, Page 7166, Year 2021
Publish dateDec 9, 2021
AuthorsChris H Hill / Lukas Pekarek / Sawsan Napthine / Anuja Kibe / Andrew E Firth / Stephen C Graham / Neva Caliskan / Ian Brierley /
PubMed AbstractProgrammed -1 ribosomal frameshifting (PRF) in cardioviruses is activated by the 2A protein, a multi-functional virulence factor that also inhibits cap-dependent translational initiation. Here we ...Programmed -1 ribosomal frameshifting (PRF) in cardioviruses is activated by the 2A protein, a multi-functional virulence factor that also inhibits cap-dependent translational initiation. Here we present the X-ray crystal structure of 2A and show that it selectively binds to a pseudoknot-like conformation of the PRF stimulatory RNA element in the viral genome. Using optical tweezers, we demonstrate that 2A stabilises this RNA element, likely explaining the increase in PRF efficiency in the presence of 2A. Next, we demonstrate a strong interaction between 2A and the small ribosomal subunit and present a cryo-EM structure of 2A bound to initiated 70S ribosomes. Multiple copies of 2A bind to the 16S rRNA where they may compete for binding with initiation and elongation factors. Together, these results define the structural basis for RNA recognition by 2A, show how 2A-mediated stabilisation of an RNA pseudoknot promotes PRF, and reveal how 2A accumulation may shut down translation during virus infection.
External linksNat Commun / PubMed:34887415 / PubMed Central
MethodsEM (single particle) / X-ray diffraction
Resolution2.62 - 2.66 Å
Structure data

12635

7nwt

EMDB-12635, PDB-7nwt:
Initiated 70S ribosome in complex with 2A protein from encephalomyocarditis virus (EMCV)
Method: EM (single particle) / Resolution: 2.66 Å

PDB-7bny:
Structure of 2A protein from encephalomyocarditis virus (EMCV)
Method: X-RAY DIFFRACTION / Resolution: 2.62 Å

Chemicals

ChemComp-SO4:
SULFATE ION

ChemComp-HOH:
WATER

ChemComp-MG:
Unknown entry

ChemComp-FME:
N-FORMYLMETHIONINE

ChemComp-ZN:
Unknown entry

Source
  • escherichia coli (E. coli)
  • mengo encephalomyocarditis virus
  • encephalomyocarditis virus
KeywordsVIRAL PROTEIN / EMCV / cardiovirus / 2A / picornavirus / frameshifting / PRF / RNA-binding protein / protein-mediated frameshifting / ribosome-binding protein / beta-shell / RIBOSOME / 70S ribosome / initiation complex

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