Lütcke H A, Chow K C, Mickel F S, Moss K A, Kern H F, Scheele G A
EMBO J. 1987 Jan;6(1):43-8. doi: 10.1002/j.1460-2075.1987.tb04716.x.
The influence of the nucleotide at position -3 relative to the AUG initiation codon on the initiation of protein synthesis was studied in two different in vitro translation systems using synthetic mRNAs. The four mRNAs, transcribed from cDNAs directed by an SP6 promoter, were identical except for mutations at nucleotide -3. In each case, translation of mRNAs produced a single protein of Mr = 12,600. Relative translational efficiencies showed a hierarchy in the reticulocyte lysate system (100, 85, 61 and 38% for A, G, U and C in position -3, respectively) but no differences in the wheat germ system. Differential mRNA degradation or polypeptide chain elongation were excluded as causes of the differences observed in translation in the reticulocyte lysate. mRNA competition increased the differences observed in translational efficiencies in reticulocyte lysate but showed no effect in wheat germ. Analysis of 61 plant and 209 animal mRNA sequences revealed qualitative and quantitative differences between the consensus sequences surrounding AUG initiation codons. Whereas the consensus sequence for animals was CACCAUG that for plants was AACAAUGGC. Both the structural and functional findings suggest that the factors which select AUG initiation codons in plants and animals differ significantly.
利用合成mRNA,在两种不同的体外翻译系统中研究了相对于AUG起始密码子的-3位核苷酸对蛋白质合成起始的影响。由SP6启动子指导从cDNA转录而来的四种mRNA,除了-3位核苷酸处的突变外,其余均相同。在每种情况下,mRNA的翻译都产生了一种分子量为12,600的单一蛋白质。相对翻译效率在网织红细胞裂解物系统中呈现出一种等级关系(-3位分别为A、G、U和C时,相对效率分别为100%、85%、61%和38%),但在小麦胚芽系统中没有差异。网织红细胞裂解物中观察到的翻译差异并非由mRNA降解差异或多肽链延伸差异所致。mRNA竞争加剧了网织红细胞裂解物中观察到的翻译效率差异,但在小麦胚芽中没有影响。对61个植物和209个动物mRNA序列的分析揭示了AUG起始密码子周围共有序列在质量和数量上的差异。动物的共有序列是CACCAUG,而植物的是AACAAUGGC。结构和功能研究结果均表明,植物和动物中选择AUG起始密码子的因子存在显著差异。