Release factor RF3 abolishes competition between release factor RF1 and ribosome recycling factor (RRF) for a ribosome binding site.
The dependence of the rate of ribosomal recycling (from initiation via protein elongation and termination, and then back to initiation) on the concentrations of release factor RF1 and the ribosome recycling factor (RRF) has been studied in vitro. High RF1 concentration was found to reduce the rate of ribosomal recycling and the extent of this reduction depended on stop codon context. The inhibitory effect of high RF1 concentrations can be reversed by a corresponding increase in RRF concentration. This indicates that RF1 and RRF have mutually exclusive and perhaps overlapping binding sites on the ribosome. Addition of release factor RF3 to the translation system abolishes the inhibitory effect of high RF1 concentration and increases the overall rate of ribosome recycling. These data can be explained by a three-step model for termination where the first step is RF1-promoted hydrolysis of peptidyl-tRNA. The second step is an intrinsically slow dissociation of RF1 which is accelerated by RF3. The third step, catalysed by RRF and elongation factor G, leads to mobility of the ribosome on mRNA allowing it to enter a further round of translation. In the absence of RF3, RF1 can re-associate rapidly with the ribosome after peptidyl-tRNA hydrolysis, preventing RRF from entering the ribosomal A-site and thereby inhibiting ribosomal recycling. The overproduction of RF1 in cells deficient in RRF or lacking RF3 has effects on growth rate predicted by the in vitro experiments.
Pavlov
M Y
MY
Department of Molecular Biology, BMC, Uppsala, S-75124, Sweden.
Freistroffer
D V
DV
Heurgué-Hamard
V
V
Buckingham
R H
RH
Ehrenberg
M
M
D001426:Bacterial Proteins / Q000378:metabolism*; D002474:Cell-Free System; D018388:Codon, Terminator; D004926:Escherichia coli / Q000254:growth & development; D029968:Escherichia coli Proteins; D008957:Models, Genetic; D008962:Models, Theoretical; D009154:Mutation; D010443:Peptide Chain Termination, Translational; D010454:Peptide Termination Factors / Q000378:metabolism*; D011485:Protein Binding; D014176:Protein Biosynthesis; D011506:Proteins; D012269:Ribosomal Proteins; D012270:Ribosomes / Q000378:metabolism*
D001426:Bacterial Proteins; D018388:Codon, Terminator; D029968:Escherichia coli Proteins; D010454:Peptide Termination Factors; D011506:Proteins; D012269:Ribosomal Proteins; C087971:peptide-chain-release factor 3; C489005:prfA protein, E coli; C489007:prfC protein, E coli; C087267:ribosome releasing factor