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TitleStructural basis for +1 ribosomal frameshifting during EF-G-catalyzed translocation.
Journal, issue, pagesNat Commun, Vol. 12, Issue 1, Page 4644, Year 2021
Publish dateJul 30, 2021
AuthorsGabriel Demo / Howard B Gamper / Anna B Loveland / Isao Masuda / Christine E Carbone / Egor Svidritskiy / Ya-Ming Hou / Andrei A Korostelev /
PubMed AbstractFrameshifting of mRNA during translation provides a strategy to expand the coding repertoire of cells and viruses. How and where in the elongation cycle +1-frameshifting occurs remains poorly ...Frameshifting of mRNA during translation provides a strategy to expand the coding repertoire of cells and viruses. How and where in the elongation cycle +1-frameshifting occurs remains poorly understood. We describe seven ~3.5-Å-resolution cryo-EM structures of 70S ribosome complexes, allowing visualization of elongation and translocation by the GTPase elongation factor G (EF-G). Four structures with a + 1-frameshifting-prone mRNA reveal that frameshifting takes place during translocation of tRNA and mRNA. Prior to EF-G binding, the pre-translocation complex features an in-frame tRNA-mRNA pairing in the A site. In the partially translocated structure with EF-G•GDPCP, the tRNA shifts to the +1-frame near the P site, rendering the freed mRNA base to bulge between the P and E sites and to stack on the 16S rRNA nucleotide G926. The ribosome remains frameshifted in the nearly post-translocation state. Our findings demonstrate that the ribosome and EF-G cooperate to induce +1 frameshifting during tRNA-mRNA translocation.
External linksNat Commun / PubMed:34330903 / PubMed Central
MethodsEM (single particle)
Resolution3.2 - 3.5 Å
Structure data

22669

7k50

EMDB-22669: Cryo-EM map of pre-translocation non-frameshifting(CCA-A) complex (Structure I)
PDB-7k50: Pre-translocation non-frameshifting(CCA-A) complex (Structure I)
Method: EM (single particle) / Resolution: 3.4 Å

22670

7k51

EMDB-22670: Cryo-EM map of mid-translocated non-frameshifting(CCA-A) complex with EF-G and GDPCP (Structure II)
PDB-7k51: Mid-translocated non-frameshifting(CCA-A) complex with EF-G and GDPCP (Structure II)
Method: EM (single particle) / Resolution: 3.5 Å

22671

7k52

EMDB-22671: Cryo-EM map of near post-translocated non-frameshifting(CCA-A) complex with EF-G and GDPCP (Structure III)
PDB-7k52: Near post-translocated non-frameshifting(CCA-A) complex with EF-G and GDPCP (Structure III)
Method: EM (single particle) / Resolution: 3.4 Å

22672

7k53

EMDB-22672: Cryo-EM map of pre-translocation +1-frameshifting(CCC-A) complex (Structure I-FS)
PDB-7k53: Pre-translocation +1-frameshifting(CCC-A) complex (Structure I-FS)
Method: EM (single particle) / Resolution: 3.2 Å

22673

7k54

EMDB-22673: Cryo-EM of mid-translocated +1-frameshifting(CCC-A) complex with EF-G and GDPCP (Structure II-FS)
PDB-7k54: Mid-translocated +1-frameshifting(CCC-A) complex with EF-G and GDPCP (Structure II-FS)
Method: EM (single particle) / Resolution: 3.2 Å

22674

7k55

EMDB-22674: Cryo-EM map of near post-translocated +1-frameshifting(CCC-A) complex with EF-G and GDPCP (Structure III-FS)
PDB-7k55: Near post-translocated +1-frameshifting(CCC-A) complex with EF-G and GDPCP (Structure III-FS)
Method: EM (single particle) / Resolution: 3.3 Å

23528

7lv0

EMDB-23528: Cryo-EM of pre-translocation rotated ribosome +1-frameshifting(CCC-A) complex (Structure Irot-FS)
PDB-7lv0: Pre-translocation rotated ribosome +1-frameshifting(CCC-A) complex (Structure Irot-FS)
Method: EM (single particle) / Resolution: 3.2 Å

Chemicals

ChemComp-FME:
N-FORMYLMETHIONINE

ChemComp-PRO:
PROLINE

ChemComp-GCP:
PHOSPHOMETHYLPHOSPHONIC ACID GUANYLATE ESTER / GMP-PCP, energy-carrying molecule analogue*YM

ChemComp-MG:
Unknown entry

Source
  • escherichia coli (strain k12) (bacteria)
  • escherichia coli k-12 (bacteria)
  • escherichia coli (E. coli)
KeywordsRIBOSOME / tRNA / TRANSLATION / EFG / EF-G

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