2 The Pathway and Mechanism of Eukaryotic Protein Synthesis
2 The Pathway and Mechanism of Eukaryotic Protein Synthesis
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DOI：
10.1101/087969458.30.31
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发表时间：
1996
期刊：
影响因子：
--
通讯作者：
W. Merrick;J. Hershey
中科院分区：
文献类型：
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作者：
W. Merrick;J. Hershey
了解蛋白质合成的详细机制对于理解翻译控制是必不可少的。我们主要关注的是翻译装置中的各种大分子如何相互作用以促进真核细胞中蛋白质合成的过程。整个过程可方便地分为三个阶段：起始、延伸和终止。本章重点介绍可溶性因子，即起始因子、延伸因子和释放因子如何催化氨酰-tRNA与核糖体的顺序结合和反应，这些反应由模板信使RNA决定。关于转运RNA的结构、氨酰化和核糖体结构的细节不在本章的讨论范围之内。有关氨酰-tRNA的形成和功能的综述，参见Carter（1993）和Soll（1993）;也参见Wool（本卷）关于核糖体蛋白的章节和一本关于核糖体结构/功能的书（Nierhaus et al. 1993）。对蛋白质合成过程的了解主要来自生物化学研究，这些研究利用了来自细菌或哺乳动物细胞的放射性标记氨基酸和分级裂解物。通过纯化在试管中重建翻译所需的蛋白质和核酸来鉴定主要的大分子组分。令人惊讶的是，细菌遗传学方法仅对鉴定组成翻译装置的200多种大分子组分做出了适度的贡献。由于生物化学方法如此富有成效，随后关于这些分子如何相互作用的体外研究进展迅速。直到最近，用酵母酿酒酵母进行的遗传研究或使用重组DNA技术的实验才使研究人员能够研究这种机制。
Knowledge of the detailed mechanism of protein synthesis is essential for understanding translational controls. We are primarily concerned with how the various macromolecules of the translational apparatus interact to promote the process of protein synthesis in eukaryotic cells. The entire process is divided conveniently into three phases: initiation, elongation, and termination. This chapter focuses on how the soluble factors, namely, initiation factors, elongation factors, and release factors, catalyze the sequential binding and reaction of aminoacyl-tRNAs to ribosomes as dictated by the template messenger RNA. Details of transfer RNA structure and aminoacylation and ribosome structure lie outside the scope of this chapter. For reviews on the formation and functions of aminoacyl-tRNAs, see Carter (1993) and Soll (1993); see also the chapter by Wool (this volume) on ribosomal proteins and a book on ribosome structure/function (Nierhaus et al. 1993). Insight into the process of protein synthesis emerged primarily from biochemical studies that utilized radioactively labeled amino acids and fractionated lysates derived from either bacterial or mammalian cells. The major macromolecular components were identified by purifying proteins and nucleic acids required to reconstitute translation in the test tube. Surprisingly, bacterial genetic approaches contributed only modestly to the identification of the greater than 200 macromolecular components that comprise the translational apparatus. Because the biochemical approach was so fruitful, subsequent in vitro studies on how these molecules interact proceeded rapidly. It is only recently that genetic studies with the yeast Saccharomyces cerevisiae or experiments using recombinant DNA techniques have enabled researchers to examine the mechanism...