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TitleStructure of the ribosome post-recycling complex probed by chemical cross-linking and mass spectrometry.
Journal, issue, pagesNat Commun, Vol. 7, Page 13248, Year 2016
Publish dateNov 8, 2016
AuthorsKristin Kiosze-Becker / Alessandro Ori / Milan Gerovac / André Heuer / Elina Nürenberg-Goloub / Umar Jan Rashid / Thomas Becker / Roland Beckmann / Martin Beck / Robert Tampé /
PubMed AbstractRibosome recycling orchestrated by the ATP binding cassette (ABC) protein ABCE1 can be considered as the final-or the first-step within the cyclic process of protein synthesis, connecting translation ...Ribosome recycling orchestrated by the ATP binding cassette (ABC) protein ABCE1 can be considered as the final-or the first-step within the cyclic process of protein synthesis, connecting translation termination and mRNA surveillance with re-initiation. An ATP-dependent tweezer-like motion of the nucleotide-binding domains in ABCE1 transfers mechanical energy to the ribosome and tears the ribosome subunits apart. The post-recycling complex (PRC) then re-initiates mRNA translation. Here, we probed the so far unknown architecture of the 1-MDa PRC (40S/30S·ABCE1) by chemical cross-linking and mass spectrometry (XL-MS). Our study reveals ABCE1 bound to the translational factor-binding (GTPase) site with multiple cross-link contacts of the helix-loop-helix motif to the S24e ribosomal protein. Cross-linking of the FeS cluster domain to the ribosomal protein S12 substantiates an extreme lever-arm movement of the FeS cluster domain during ribosome recycling. We were thus able to reconstitute and structurally analyse a key complex in the translational cycle, resembling the link between translation initiation and ribosome recycling.
External linksNat Commun / PubMed:27824037 / PubMed Central
MethodsEM (single particle)
Resolution17.0 Å
Structure data

4113

5lw7

EMDB-4113: 30S-ABCE1 Post-Recycling Complex
PDB-5lw7: S. solfataricus ABCE1 post-splitting state
Method: EM (single particle) / Resolution: 17.0 Å

Chemicals

ChemComp-SF4:
IRON/SULFUR CLUSTER

Source
  • Sulfolobus solfataricus (archaea)
  • pyrococcus abyssi (strain ge5 / orsay) (archaea)
KeywordsRIBOSOME / ABCE1 / recycling / 30S

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