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PMID: 9233821 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Release factor RF3 in E.coli accelerates the dissociation of release factors RF1 and RF2 from the ribosome in a GTP-dependent manner.

The EMBO journal ·Vol. 16 ·No. 13 ·1997-07-01 ·Pages 4126-33

Freistroffer DV, Pavlov MY, MacDougall J, Buckingham RH, Ehrenberg M

Abstract

Ribosomes complexed with synthetic mRNA and peptidyl-tRNA, ready for peptide release, were purified by gel filtration and used to study the function of release factor RF3 and guanine nucleotides in the termination of protein synthesis. The peptide-releasing activity of RF1 and RF2 in limiting concentrations was stimulated by the addition of RF3 and GTP, stimulated, though to a lesser extent, by RF3 and a non-hydrolysable GTP analogue, and inhibited by RF3 and GDP or RF3 without guanine nucleotide. With short incubation times allowing only a single catalytic cycle of RF1 or RF2, peptide release activity was independent of RF3 and guanine nucleotide. RF3 hydrolysis of GTP to GDP + P(i) was dependent only on ribosomes and not on RF1 or RF2. RF3 affected neither the rate of association of RF1 and RF2 with the ribosome nor the catalytic rate of peptide release. A model is proposed which explains how RF3 recycles RF1 and RF2 by displacing the factors from the ribosome after the release of peptide.

MeSH Terms
Bacterial Proteins/metabolism Codon, Terminator Escherichia coli/metabolism Escherichia coli Proteins Guanosine Diphosphate/metabolism Guanosine Triphosphate/metabolism Hydrolysis Peptide Termination Factors/metabolism RNA, Transfer, Amino Acyl/metabolism Ribosomes/metabolism
Chemicals
Bacterial Proteins Codon, Terminator Escherichia coli Proteins Peptide Termination Factors RNA, Transfer, Amino Acyl peptide chain termination release factor 2 peptide-chain-release factor 3 prfA protein, E coli prfB protein, E coli prfC protein, E coli tRNA, peptidyl- Guanosine Diphosphate Guanosine Triphosphate
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Freistroffer D V
Department of Molecular Biology, Uppsala University, BMC, Sweden.
Pavlov M Y
MacDougall J
Buckingham R H
Ehrenberg M
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/16.13.4126
Published
1997-07-01
Pages
4126-33
Language
English
Region
England
NLM ID
8208664
PMCID
PMC1170035
Subset
IM
External Links
PubMed source DOI: 10.1093/emboj/16.13.4126 PMC Full Text View journal details
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