Mottagui-Tabar S, Tuite M F, Isaksson L A
Department of Biosciences, University of Kent at Canterbury, United Kingdom.
Eur J Biochem. 1998 Oct 1;257(1):249-54. doi: 10.1046/j.1432-1327.1998.2570249.x.
Translation termination in vivo was studied in the yeast Saccharomyces cerevisiae using a translation-assay system. Codon changes that were made at position -2 relative to the stop codon, gave a 3.5-fold effect on termination in a release-factor-defective (sup45) mutant strain, in line with the effect observed in a wild-type strain. The influence of the -2 codon could be correlated to the charge of the corresponding amino acid residue in the nascent peptide; an acidic residue favoring efficient termination. Thus, the C-terminal end of the nascent peptide influences translation termination both in the bacterium Escherichia coli and to a lesser extent in the yeast S. cerevisiae. However, the sensitivity to the charge of the penultimate amino acid is reversed when the E. coli and S. cerevisiae are compared. Changing - 1 (P-site) codons in yeast gave a 10-fold difference in effect on the efficiency of termination. This effect could not be related to any property of the encoded last amino acid in the nascent peptide. Iso-codons read by the same tRNA (AAA/G, GAA/G) gave similar readthrough values. Codons for glutamine (CAA/G), glutamic acid (GAA/G) and isoleucine (AUA/C) that are read by different isoaccepting tRNAs are associated with an approximately twofold difference in each case in termination efficiency. This suggests that the P-site tRNA is able to influence termination at UGAC in yeast.
利用翻译分析系统,在酿酒酵母中研究了体内翻译终止过程。相对于终止密码子在第 -2 位进行的密码子改变,在释放因子缺陷型(sup45)突变菌株中对终止产生了 3.5 倍的影响,这与在野生型菌株中观察到的影响一致。-2 密码子的影响可能与新生肽中相应氨基酸残基的电荷相关;酸性残基有利于高效终止。因此,新生肽的 C 末端在大肠杆菌和酿酒酵母中均影响翻译终止,不过在酿酒酵母中的影响程度较小。然而,当比较大肠杆菌和酿酒酵母时,对倒数第二个氨基酸电荷的敏感性是相反的。在酵母中改变 -1(P 位点)密码子对终止效率的影响有 10 倍差异。这种影响与新生肽中编码的最后一个氨基酸的任何特性均无关。由相同 tRNA(AAA/G、GAA/G)读取的同义密码子给出了相似的通读值。由不同同工受体 tRNA 读取的谷氨酰胺(CAA/G)、谷氨酸(GAA/G)和异亮氨酸(AUA/C)密码子,在每种情况下终止效率均存在约两倍的差异。这表明 P 位点 tRNA 能够影响酵母中 UGA 处的终止。