Department of Molecular Genetics, Center for RNA Biology, The Ohio State University, Columbus, Ohio 43210, USA.
Genetics. 2013 May;194(1):43-67. doi: 10.1534/genetics.112.147470.
Transfer RNAs (tRNAs) are essential for protein synthesis. In eukaryotes, tRNA biosynthesis employs a specialized RNA polymerase that generates initial transcripts that must be subsequently altered via a multitude of post-transcriptional steps before the tRNAs beome mature molecules that function in protein synthesis. Genetic, genomic, biochemical, and cell biological approaches possible in the powerful Saccharomyces cerevisiae system have led to exciting advances in our understandings of tRNA post-transcriptional processing as well as to novel insights into tRNA turnover and tRNA subcellular dynamics. tRNA processing steps include removal of transcribed leader and trailer sequences, addition of CCA to the 3' mature sequence and, for tRNA(His), addition of a 5' G. About 20% of yeast tRNAs are encoded by intron-containing genes. The three-step splicing process to remove the introns surprisingly occurs in the cytoplasm in yeast and each of the splicing enzymes appears to moonlight in functions in addition to tRNA splicing. There are 25 different nucleoside modifications that are added post-transcriptionally, creating tRNAs in which ∼15% of the residues are nucleosides other than A, G, U, or C. These modified nucleosides serve numerous important functions including tRNA discrimination, translation fidelity, and tRNA quality control. Mature tRNAs are very stable, but nevertheless yeast cells possess multiple pathways to degrade inappropriately processed or folded tRNAs. Mature tRNAs are also dynamic in cells, moving from the cytoplasm to the nucleus and back again to the cytoplasm; the mechanism and function of this retrograde process is poorly understood. Here, the state of knowledge for tRNA post-transcriptional processing, turnover, and subcellular dynamics is addressed, highlighting the questions that remain.
转移 RNA(tRNA)对于蛋白质合成至关重要。在真核生物中,tRNA 生物合成采用专门的 RNA 聚合酶,该酶生成初始转录物,然后必须通过多种转录后步骤进行改变,然后 tRNA 才能成为在蛋白质合成中起作用的成熟分子。在强大的酿酒酵母系统中可以采用遗传、基因组、生化和细胞生物学方法,这使得我们对 tRNA 转录后加工的理解取得了令人兴奋的进展,并对 tRNA 周转和 tRNA 亚细胞动力学有了新的认识。tRNA 加工步骤包括去除转录的前导序列和尾随序列,在 3'成熟序列上添加 CCA,对于 tRNA(His),在 5'添加 G。约 20%的酵母 tRNA 由内含子基因编码。令人惊讶的是,在酵母中,三步剪接过程去除内含子发生在细胞质中,除了 tRNA 剪接外,每个剪接酶似乎都有兼职功能。有 25 种不同的核苷修饰物是转录后添加的,这使得 tRNA 中约 15%的残基不是 A、G、U 或 C。这些修饰核苷具有许多重要功能,包括 tRNA 鉴别、翻译保真度和 tRNA 质量控制。成熟的 tRNA 非常稳定,但尽管如此,酵母细胞仍具有多种途径来降解处理不当或折叠的 tRNA。成熟的 tRNA 在细胞中也是动态的,从细胞质到细胞核再回到细胞质;这个逆行过程的机制和功能知之甚少。本文讨论了 tRNA 转录后加工、周转和亚细胞动力学的现状,强调了仍然存在的问题。