Dynamics of release factor recycling during translation termination in bacteria.


Journal

Nucleic acids research
ISSN: 1362-4962
Titre abrégé: Nucleic Acids Res
Pays: England
ID NLM: 0411011

Informations de publication

Date de publication:
23 06 2023
Historique:
accepted: 09 04 2023
revised: 05 04 2023
received: 02 07 2022
medline: 26 6 2023
pubmed: 27 4 2023
entrez: 27 4 2023
Statut: ppublish

Résumé

In bacteria, release of newly synthesized proteins from ribosomes during translation termination is catalyzed by class-I release factors (RFs) RF1 or RF2, reading UAA and UAG or UAA and UGA codons, respectively. Class-I RFs are recycled from the post-termination ribosome by a class-II RF, the GTPase RF3, which accelerates ribosome intersubunit rotation and class-I RF dissociation. How conformational states of the ribosome are coupled to the binding and dissociation of the RFs remains unclear and the importance of ribosome-catalyzed guanine nucleotide exchange on RF3 for RF3 recycling in vivo has been disputed. Here, we profile these molecular events using a single-molecule fluorescence assay to clarify the timings of RF3 binding and ribosome intersubunit rotation that trigger class-I RF dissociation, GTP hydrolysis, and RF3 dissociation. These findings in conjunction with quantitative modeling of intracellular termination flows reveal rapid ribosome-dependent guanine nucleotide exchange to be crucial for RF3 action in vivo.

Identifiants

pubmed: 37102635
pii: 7145697
doi: 10.1093/nar/gkad286
pmc: PMC10287982
doi:

Substances chimiques

Guanosine Triphosphate 86-01-1
Peptide Termination Factors 0

Types de publication

Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

5774-5790

Subventions

Organisme : NIGMS NIH HHS
ID : R01 GM113078
Pays : United States
Organisme : NIGMS NIH HHS
ID : T32 GM008294
Pays : United States
Organisme : NIGMS NIH HHS
ID : R01 GM051266
Pays : United States

Informations de copyright

© The Author(s) 2023. Published by Oxford University Press on behalf of Nucleic Acids Research.

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Auteurs

Arjun Prabhakar (A)

Department of Structural Biology, Stanford University School of Medicine, Stanford, CA 94305-5126, USA.
Program in Biophysics, Stanford University, Stanford, CA 94305-5126, USA.

Michael Y Pavlov (MY)

Department of Cell and Molecular Biology, Biomedical Center, Box 596, Uppsala University, Uppsala, Sweden.

Jingji Zhang (J)

Department of Structural Biology, Stanford University School of Medicine, Stanford, CA 94305-5126, USA.

Gabriele Indrisiunaite (G)

Department of Cell and Molecular Biology, Biomedical Center, Box 596, Uppsala University, Uppsala, Sweden.

Jinfan Wang (J)

Department of Structural Biology, Stanford University School of Medicine, Stanford, CA 94305-5126, USA.

Michael R Lawson (MR)

Department of Structural Biology, Stanford University School of Medicine, Stanford, CA 94305-5126, USA.

Måns Ehrenberg (M)

Department of Cell and Molecular Biology, Biomedical Center, Box 596, Uppsala University, Uppsala, Sweden.

Joseph D Puglisi (JD)

Department of Structural Biology, Stanford University School of Medicine, Stanford, CA 94305-5126, USA.

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