Functional interaction between release factor one and P-site peptidyl-tRNA on the ribosome.
Translation termination at UAG is influenced by the nature of the 5' flanking codon in Escherichia coli. Readthrough of the stop codon is always higher in a strain with mutant (prfA1) as compared to wild-type (prfA+) release factor one (RF1). Isocodons, which differ in the last base and are decoded by the same tRNA species, affect termination at UAG differently in strains with mutant or wild-type RF1. No general preference of the last codon base to favour readthrough or termination can be found. The data suggest that RF1 is sensitive to the nature of the wobble base anticodon-codon interaction at the ribosomal peptidyl-tRNA binding site (P-site). For some isoaccepting P-site tRNAs (tRNA3(Pro) versus tRNA2(Pro), tRNA4(Thr) versus tRNA1,3Thr) the effect is different on mutant and wild-type RF1, suggesting an interaction between RF1 at the aminoacyl-tRNA acceptor site (A-site) and the P-site tRNA itself. The glycine codons GGA (tRNA2(Gly)) and GGG (tRNA2,3(Gly)) at the ribosomal P-site are associated with an almost threefold higher readthrough of UAG than any of the other 42 codons tested, including the glycine codons GGU/C, in a strain with wild-type RF1. This differential response to the glycine codons is lost in the strain with the mutant form of RF1 since readthrough is increased to a similar high level for all four glycine codons. High alpha-helix propensity of the last amino acid residue at the C-terminal end of the nascent peptide is correlated with an increased termination at UAG. The effect is stronger on mutant compared to wild-type RF1. The data suggest that RF1-mediated termination at UAG is sensitive to the nature of the codon-anticodon interaction of the wobble base, the last amino acid residue of the nascent peptide chain, and the tRNA at the ribosomal P-site.
Zhang
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S
Department of Microbiology Stockholm University, Sweden.
Rydén-Aulin
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Isaksson
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D000596:Amino Acids / Q000235:genetics; D001483:Base Sequence; D003062:Codon / Q000235:genetics; D018388:Codon, Terminator / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D029968:Escherichia coli Proteins; D005998:Glycine / Q000235:genetics; D008969:Molecular Sequence Data; D009154:Mutation; D010443:Peptide Chain Termination, Translational / Q000235:genetics*; D010454:Peptide Termination Factors / Q000235:genetics / Q000502:physiology*; D012346:RNA, Transfer, Amino Acyl / Q000378:metabolism*; D011993:Recombinant Fusion Proteins; D012270:Ribosomes / Q000378:metabolism*; D013205:Staphylococcal Protein A / Q000235:genetics
D000596:Amino Acids; D003062:Codon; D018388:Codon, Terminator; D029968:Escherichia coli Proteins; D010454:Peptide Termination Factors; D012346:RNA, Transfer, Amino Acyl; D011993:Recombinant Fusion Proteins; D013205:Staphylococcal Protein A; C489005:prfA protein, E coli; D005998:Glycine