Spontaneous ribosomal translocation of mRNA and tRNAs into a chimeric hybrid state

Zhou, Jie; Lancaster, Laura; Donohue, John Paul; Noller, Harry F.

doi:10.1073/pnas.1901310116

Title: Spontaneous ribosomal translocation of mRNA and tRNAs into a chimeric hybrid state

Journal Article · Mon Apr 01 00:00:00 EDT 2019 · Proceedings of the National Academy of Sciences of the United States of America

DOI:https://doi.org/10.1073/pnas.1901310116· OSTI ID:1578229

Zhou, Jie ^[1]; Lancaster, Laura ^[1]; Donohue, John Paul ^[1];

^[1]

Univ. of California, Santa Cruz, CA (United States)

The elongation factor G (EF-G)–catalyzed translocation of mRNA and tRNA through the ribosome is essential for vacating the ribosomal A site for the next incoming aminoacyl-tRNA, while precisely maintaining the translational reading frame. Here, the 3.2-Å crystal structure of a ribosome translocation intermediate complex containing mRNA and two tRNAs, formed in the absence of EF-G or GTP, provides insight into the respective roles of EF-G and the ribosome in translocation. Unexpectedly, the head domain of the 30S subunit is rotated by 21°, creating a ribosomal conformation closely resembling the two-tRNA chimeric hybrid state that was previously observed only in the presence of bound EF-G. The two tRNAs have moved spontaneously from their A/A and P/P binding states into ap/P and pe/E states, in which their anticodon loops are bound between the 30S body domain and its rotated head domain, while their acceptor ends have moved fully into the 50S P and E sites, respectively. Remarkably, the A-site tRNA translocates fully into the classical P-site position. Furthermore, although the mRNA also undergoes movement, codon–anticodon interaction is disrupted in the absence of EF-G, resulting in slippage of the translational reading frame. We conclude that, although movement of both tRNAs and mRNA (along with rotation of the 30S head domain) can occur in the absence of EF-G and GTP, EF-G is essential for enforcing coupled movement of the tRNAs and their mRNA codons to maintain the reading frame.

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Research Organization:: Argonne National Laboratory (ANL), Argonne, IL (United States). Advanced Photon Source (APS)

Sponsoring Organization:: National Institutes of Health (NIH); Robert L. Sinsheimer Chair

Grant/Contract Number:: R35-GM118156

OSTI ID:: 1578229

Journal Information:: Proceedings of the National Academy of Sciences of the United States of America, Vol. 116, Issue 16; ISSN 0027-8424

Publisher:: National Academy of SciencesCopyright Statement

Country of Publication:: United States

Language:: ENGLISH

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Cited by: 27 works

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Title: Spontaneous ribosomal translocation of mRNA and tRNAs into a chimeric hybrid state

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