Here, we visualize by cryo-electron microscopy (cryo-EM) how Drg1 is recruited to the pre-ribosome to extract Rlp24. Rlp24 is captured by the Drg1 N domains and AAA domains and extracted by hand-over-hand translocation. Our data uncover orchestrated conformational changes of the AAA-ATPase during substrate translocation and provide a structural basis for the substrate-processing mechanism.
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