Blake K R, Murakami A, Miller P S
Biochemistry. 1985 Oct 22;24(22):6132-8. doi: 10.1021/bi00343a015.
Oligodeoxyribonucleotides 8-12 nucleotides in length whose sequences are complementary to the 5' end, the initiation codon regions, or the coding regions of rabbit globin mRNA were tested for their ability to inhibit translation in a rabbit reticulocyte lysate and in a wheat germ extract. The oligomers interact specifically with their target mRNAs as shown by their ability to serve as primers with reverse transcriptase. In the reticulocyte lysate, oligomers complementary to the 5' end or the initiation codon regions inhibit translation of both alpha- and beta-globin mRNA, whereas oligomers complementary to the coding regions have little or no effect. This suggests that reticulocyte ribosomes are able to displace the oligomers from the mRNA during the elongation but not the initiation step of translation. In the wheat germ system, translation was effectively inhibited by all oligomers regardless of their binding site on the message. In contrast to their behavior in the reticulocyte system, the oligomers inhibited alpha- and beta-globin synthesis in a specific manner. This observation suggests that control of alpha- and beta-globin mRNA translation is coordinated in the reticulocyte lysate system but not in the wheat germ extract. The results of our studies indicate that oligodeoxyribonucleotides may be useful probes for studying control of mRNA translation in cell-free systems.
对长度为8 - 12个核苷酸的寡脱氧核糖核苷酸进行了测试，这些寡核苷酸的序列与兔珠蛋白mRNA的5'端、起始密码子区域或编码区域互补，以检测它们在兔网织红细胞裂解物和小麦胚芽提取物中抑制翻译的能力。这些寡聚物能够作为逆转录酶的引物，表明它们与目标mRNA发生特异性相互作用。在网织红细胞裂解物中，与5'端或起始密码子区域互补的寡聚物抑制α - 和β - 珠蛋白mRNA的翻译，而与编码区域互补的寡聚物几乎没有作用或没有作用。这表明网织红细胞核糖体在翻译的延伸步骤而非起始步骤中能够将寡聚物从mRNA上置换下来。在小麦胚芽系统中，所有寡聚物均能有效抑制翻译，无论它们在mRNA上的结合位点如何。与它们在网织红细胞系统中的行为相反，这些寡聚物以一种特异性方式抑制α - 和β - 珠蛋白的合成。这一观察结果表明，在网织红细胞裂解物系统中α - 和β - 珠蛋白mRNA翻译的调控是协调的，而在小麦胚芽提取物中则不然。我们的研究结果表明，寡脱氧核糖核苷酸可能是用于研究无细胞系统中mRNA翻译调控的有用探针。
