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TitleVisualization of translation termination intermediates trapped by the Apidaecin 137 peptide during RF3-mediated recycling of RF1.
Journal, issue, pagesNat Commun, Vol. 9, Issue 1, Page 3053, Year 2018
Publish dateAug 3, 2018
AuthorsMichael Graf / Paul Huter / Cristina Maracci / Miroslav Peterek / Marina V Rodnina / Daniel N Wilson /
PubMed AbstractDuring translation termination in bacteria, the release factors RF1 and RF2 are recycled from the ribosome by RF3. While high-resolution structures of the individual termination factors on the ...During translation termination in bacteria, the release factors RF1 and RF2 are recycled from the ribosome by RF3. While high-resolution structures of the individual termination factors on the ribosome exist, direct structural insight into how RF3 mediates dissociation of the decoding RFs has been lacking. Here we have used the Apidaecin 137 peptide to trap RF1 together with RF3 on the ribosome and visualize an ensemble of termination intermediates using cryo-electron microscopy. Binding of RF3 to the ribosome induces small subunit (SSU) rotation and swivelling of the head, yielding intermediate states with shifted P-site tRNAs and RF1 conformations. RF3 does not directly eject RF1 from the ribosome, but rather induces full rotation of the SSU that indirectly dislodges RF1 from its binding site. SSU rotation is coupled to the accommodation of the GTPase domain of RF3 on the large subunit (LSU), thereby promoting GTP hydrolysis and dissociation of RF3 from the ribosome.
External linksNat Commun / PubMed:30076302 / PubMed Central
MethodsEM (single particle)
Resolution3.8 - 4.4 Å
Structure data

0076

6gwt

EMDB-0076, PDB-6gwt:
Cryo-EM structure of an E. coli 70S ribosome in complex with RF3-GDPCP, RF1(GAQ) and Pint-tRNA (State I)
Method: EM (single particle) / Resolution: 3.8 Å

0080

6gxm

EMDB-0080, PDB-6gxm:
Cryo-EM structure of an E. coli 70S ribosome in complex with RF3-GDPCP, RF1(GAQ) and Pint-tRNA (State II)
Method: EM (single particle) / Resolution: 3.8 Å

0081

6gxn

EMDB-0081, PDB-6gxn:
Cryo-EM structure of an E. coli 70S ribosome in complex with RF3-GDPCP, RF1(GAQ) and Pint-tRNA (State III)
Method: EM (single particle) / Resolution: 3.9 Å

0082

6gxo

EMDB-0082, PDB-6gxo:
Cryo-EM structure of a rotated E. coli 70S ribosome in complex with RF3-GDPCP, RF1(GAQ) and P/E-tRNA (State IV)
Method: EM (single particle) / Resolution: 3.9 Å

0083

6gxp

EMDB-0083, PDB-6gxp:
Cryo-EM structure of a rotated E. coli 70S ribosome in complex with RF3-GDPCP(RF3-only)
Method: EM (single particle) / Resolution: 4.4 Å

Chemicals

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

Source
  • escherichia coli (E. coli)
  • apis mellifera (honey bee)
KeywordsRIBOSOME / single particle cryo-EM / GTPase / release factor RF1 / RF2 / RF3 / subunit rotation / translation termination

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