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Structure of the ribosome post-recycling complex probed by chemical cross-linking and mass spectrometry.


ABSTRACT: 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.

SUBMITTER: Kiosze-Becker K 

PROVIDER: S-EPMC5105147 | biostudies-literature | 2016 Nov

REPOSITORIES: biostudies-literature

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Structure of the ribosome post-recycling complex probed by chemical cross-linking and mass spectrometry.

Kiosze-Becker Kristin K   Ori Alessandro A   Gerovac Milan M   Heuer André A   Nürenberg-Goloub Elina E   Rashid Umar Jan UJ   Becker Thomas T   Beckmann Roland R   Beck Martin M   Tampé Robert R  

Nature communications 20161108


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 probe  ...[more]

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