Effect of spermine on the efficiency and fidelity of the codon-specific binding of tRNA to the ribosomes.
Binding of the yeast Tyr-tRNA and Phe-tRNA to the A site, and the binding of their acetyl derivatives to the P site of poly(U11,A)-programmed Escherichia coli ribosomes was studied. Spermine stimulated the rate of binding of both tRNAs at least threefold, enabling more than 90% final saturation of both ribosomal binding sites. The effect is observed when the tRNAs, but not ribosomes or poly(U11,A), are preincubated with polyamine. Regardless of the binding site, optimal saturation was reached at spermine/tRNA molar ratios of 3 for tRNA(Phe) and 5 for tRNA(Tyr). The same low spermine/tRNA ratios were previously reported to stabilize the conformation of these tRNAs in solution. On the other hand, the messenger-free, EF-Tu- and EF-G-dependent polymerization of lysine from E. coli Lys-tRNA is drastically reduced, while the poly(A)-directed polymerization is stimulated by spermine through a wide range of Mg2+ concentrations. Misreading of UUU codons as isoleucine, assayed by the A-site binding of E. coli Ile-tRNA, is also inhibited by spermine. All these results demonstrate that spermine increases the efficiency and accuracy of a series of macromolecular interactions leading to the correct incorporation of an amino acid into protein, at the same time preventing some unspecific or erroneous interactions. From the analogy with its known structural effects, it can be inferred that spermine does so by conferring on the tRNA a specific biologically functional conformation.
Naranda
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T
Department of Chemistry, Faculty of Science, University of Zagreb, Jugoslavija.
Kućan
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D001665:Binding Sites; D003062:Codon / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism; D011072:Poly U / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism*; D012335:RNA, Ribosomal / Q000378:metabolism*; D012343:RNA, Transfer / Q000378:metabolism*; D012360:RNA, Transfer, Phe / Q000378:metabolism; D012365:RNA, Transfer, Tyr / Q000378:metabolism; D013096:Spermine / Q000494:pharmacology*; D013997:Time Factors; D015003:Yeasts / Q000378:metabolism
D003062:Codon; D012333:RNA, Messenger; D012335:RNA, Ribosomal; D012360:RNA, Transfer, Phe; D012365:RNA, Transfer, Tyr; D011072:Poly U; D013096:Spermine; D012343:RNA, Transfer