Show simple item record

dc.contributor.authorCarbone, Christine E.
dc.contributor.authorLoveland, Anna B.
dc.contributor.authorGamper, Jr., Howard
dc.contributor.authorHou, Ya-Ming
dc.contributor.authorDemo, Gabriel
dc.contributor.authorKorostelev, Andrei A.
dc.date2022-08-11T08:08:27.000
dc.date.accessioned2022-08-23T15:55:46Z
dc.date.available2022-08-23T15:55:46Z
dc.date.issued2021-05-31
dc.date.submitted2021-07-14
dc.identifier.citation<p>bioRxiv 2021.05.31.446434; doi: https://doi.org/10.1101/2021.05.31.446434. <a href="https://doi.org/10.1101/2021.05.31.446434" target="_blank" title="view preprint in biorxiv">Link to preprint on bioRxiv.</a></p>
dc.identifier.doi10.1101/2021.05.31.446434
dc.identifier.urihttp://hdl.handle.net/20.500.14038/29827
dc.description<p>This article is a preprint. Preprints are preliminary reports of work that have not been certified by peer review.</p>
dc.description.abstractDuring translation, a conserved GTPase elongation factor—EF-G in bacteria or eEF2 in eukaryotes—translocates tRNA and mRNA through the ribosome. EF-G has been proposed to act as a flexible motor that propels tRNA and mRNA movement, as a rigid pawl that biases unidirectional translocation resulting from ribosome rearrangements, or by various combinations of motor- and pawl-like mechanisms. Using time-resolved cryo-EM, we visualized GTP-catalyzed translocation without inhibitors, capturing elusive structures of ribosome•EF-G intermediates at near-atomic resolution. Prior to translocation, EF-G binds near peptidyl-tRNA, while the rotated 30S subunit stabilizes the EF-G GTPase center. Reverse 30S rotation releases Pi and translocates peptidyl-tRNA and EF-G by ∼20 Å. An additional 4-Å translocation initiates EF-G dissociation from a transient ribosome state with highly swiveled 30S head. The structures visualize how nearly rigid EF-G rectifies inherent and spontaneous ribosomal dynamics into tRNA-mRNA translocation, whereas GTP hydrolysis and Pi release drive EF-G dissociation.
dc.language.isoen_US
dc.relationNow published in Nature Communications doi: 10.1038/s41467-021-27415-0
dc.rightsThe copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/4.0/
dc.subjectMolecular Biology
dc.subjectcryo-EM
dc.subjecttRNA
dc.subjectmRNA
dc.subjectInvestigative Techniques
dc.subjectMolecular Biology
dc.subjectNucleic Acids, Nucleotides, and Nucleosides
dc.titleTime-resolved cryo-EM visualizes ribosomal translocation with EF-G and GTP [preprint]
dc.typePreprint
dc.source.journaltitlebioRxiv
dc.identifier.legacyfulltexthttps://escholarship.umassmed.edu/cgi/viewcontent.cgi?article=3060&amp;context=faculty_pubs&amp;unstamped=1
dc.identifier.legacycoverpagehttps://escholarship.umassmed.edu/faculty_pubs/2034
dc.identifier.contextkey23822551
refterms.dateFOA2022-08-23T15:55:46Z
html.description.abstract<p><p id="x-x-x-p-2">During translation, a conserved GTPase elongation factor—EF-G in bacteria or eEF2 in eukaryotes—translocates tRNA and mRNA through the ribosome. EF-G has been proposed to act as a flexible motor that propels tRNA and mRNA movement, as a rigid pawl that biases unidirectional translocation resulting from ribosome rearrangements, or by various combinations of motor- and pawl-like mechanisms. Using time-resolved cryo-EM, we visualized GTP-catalyzed translocation without inhibitors, capturing elusive structures of ribosome•EF-G intermediates at near-atomic resolution. Prior to translocation, EF-G binds near peptidyl-tRNA, while the rotated 30S subunit stabilizes the EF-G GTPase center. Reverse 30S rotation releases Pi and translocates peptidyl-tRNA and EF-G by ∼20 Å. An additional 4-Å translocation initiates EF-G dissociation from a transient ribosome state with highly swiveled 30S head. The structures visualize how nearly rigid EF-G rectifies inherent and spontaneous ribosomal dynamics into tRNA-mRNA translocation, whereas GTP hydrolysis and Pi release drive EF-G dissociation.</p>
dc.identifier.submissionpathfaculty_pubs/2034
dc.contributor.departmentGraduate School of Biomedical Sciences
dc.contributor.departmentDepartment of Biochemistry and Molecular Pharmacology
dc.contributor.departmentRNA Therapeutics Institute


Files in this item

Thumbnail
Name:
2021.05.31.446434v1.full.pdf
Size:
5.893Mb
Format:
PDF

This item appears in the following Collection(s)

Show simple item record

The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
Except where otherwise noted, this item's license is described as The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.