Frameshifting at the internal stop codon within the mRNA for bacterial release factor-2 on eukaryotic ribosomes.
A translational frameshift is necessary in the synthesis of Escherichia coli release factor 2 (RF-2) to bypass an in-frame termination codon within the coding sequence. High-efficiency frameshifting around this codon can occur on eukaryotic ribosomes as well as prokaryotic ribosomes. This was determined from the relative efficiency of translation of RF-2 RNA compared with that for the other release factor RF-1, which lacks the in-frame premature stop codon. Since the termination product is unstable an absolute measure of the efficiency of frameshifting has not been possible. A gene fusion between trpE and RF-2 was carried out to give a stable termination product as well as the frameshift product, thereby allowing a direct determination of frameshifting efficiency. The extension of RF-2 RNA near its start codon with a fragment of the trpE gene, while still allowing high efficiency frameshifting on prokaryotic ribosomes, surprisingly gives a different estimate of frameshifting on the eukaryotic ribosomes than that obtained with RF-2 RNA alone. This paradox may be explained by long distance context effects on translation rates in the frameshift region created by the trpE sequences in the gene fusion, and may reflect that pausing and translation rate are fundamental factors in determining the efficiency of frameshifting.
Donly
C
C
Department of Biochemistry, University of Otago, Dunedin, New Zealand.
Williams
J
J
Richardson
C
C
Tate
W
W
D001483:Base Sequence; D003001:Cloning, Molecular; D003062:Codon; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D016368:Frameshift Mutation; D007700:Kinetics; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D010454:Peptide Termination Factors / Q000096:biosynthesis / Q000235:genetics*; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D012270:Ribosomes / Q000378:metabolism*
D003062:Codon; D015345:Oligonucleotide Probes; D010454:Peptide Termination Factors; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; C041460:peptide chain termination release factor 2