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Structure paper

TitleStructural basis of ribosomal frameshifting during translation of the SARS-CoV-2 RNA genome.
Journal, issue, pagesScience, Vol. 372, Issue 6548, Page 1306-1313, Year 2021
Publish dateJun 18, 2021
AuthorsPramod R Bhatt / Alain Scaiola / Gary Loughran / Marc Leibundgut / Annika Kratzel / Romane Meurs / René Dreos / Kate M O'Connor / Angus McMillan / Jeffrey W Bode / Volker Thiel / David Gatfield / John F Atkins / Nenad Ban /
PubMed AbstractProgrammed ribosomal frameshifting is a key event during translation of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA genome that allows synthesis of the viral RNA-dependent ...Programmed ribosomal frameshifting is a key event during translation of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA genome that allows synthesis of the viral RNA-dependent RNA polymerase and downstream proteins. Here, we present the cryo-electron microscopy structure of a translating mammalian ribosome primed for frameshifting on the viral RNA. The viral RNA adopts a pseudoknot structure that lodges at the entry to the ribosomal messenger RNA (mRNA) channel to generate tension in the mRNA and promote frameshifting, whereas the nascent viral polyprotein forms distinct interactions with the ribosomal tunnel. Biochemical experiments validate the structural observations and reveal mechanistic and regulatory features that influence frameshifting efficiency. Finally, we compare compounds previously shown to reduce frameshifting with respect to their ability to inhibit SARS-CoV-2 replication, establishing coronavirus frameshifting as a target for antiviral intervention.
External linksScience / PubMed:34029205 / PubMed Central
MethodsEM (single particle)
Resolution2.2 - 6.0 Å
Structure data

12756

7o7y

EMDB-12756, PDB-7o7y:
Rabbit 80S ribosome stalled close to the mutated SARS-CoV-2 slippery site by a pseudoknot (high resolution)
Method: EM (single particle) / Resolution: 2.2 Å

12757

7o7z

EMDB-12757, PDB-7o7z:
Rabbit 80S ribosome stalled close to the mutated SARS-CoV-2 slippery site by a pseudoknot (classified for pseudoknot)
Method: EM (single particle) / Resolution: 2.4 Å

12758

7o80

EMDB-12758, PDB-7o80:
Rabbit 80S ribosome in complex with eRF1 and ABCE1 stalled at the STOP codon in the mutated SARS-CoV-2 slippery site
Method: EM (single particle) / Resolution: 2.9 Å

12759

7o81

EMDB-12759, PDB-7o81:
Rabbit 80S ribosome colliding in another ribosome stalled by the SARS-CoV-2 pseudoknot
Method: EM (single particle) / Resolution: 3.1 Å

EMDB-12760:
Disome of rabbit ribosomes with the leading one stalled by the SARS-CoV-2 pseudoknot
Method: EM (single particle) / Resolution: 6.0 Å

Chemicals

ChemComp-SPD:
SPERMIDINE / Spermidine

ChemComp-SPM:
SPERMINE / Spermine

ChemComp-MG:
Unknown entry

ChemComp-UNX:
Unknown entry

ChemComp-ZN:
Unknown entry

ChemComp-HOH:
WATER / Water

ChemComp-GTP:
GUANOSINE-5'-TRIPHOSPHATE / GTP, energy-carrying molecule*YM / Guanosine triphosphate

ChemComp-SF4:
IRON/SULFUR CLUSTER / Iron–sulfur cluster

Source
  • oryctolagus cuniculus (rabbit)
  • severe acute respiratory syndrome coronavirus 2
  • bos taurus (cattle)
KeywordsRIBOSOME / Frameshift / virus / pseudoknot

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