Hossain Munshi Azad, Johnson Tracy L
Molecular Biology Section, Division of Biological Sciences, University of California, San Diego, La Jolla, CA, USA.
Methods Mol Biol. 2014;1126:285-98. doi: 10.1007/978-1-62703-980-2_21.
Pre-mRNA splicing is a critical step in eukaryotic gene expression, which involves removal of noncoding intron sequences from pre-mRNA and ligation of the remaining exon sequences to make a mature message. Splicing is carried out by a large ribonucleoprotein complex called the spliceosome. Since the first description of the pre-mRNA splicing reaction in the 1970s, elegant genetic and biochemical studies have revealed that the enzyme that catalyzes the reaction, the spliceosome, is an exquisitely dynamic macromolecular machine, and its RNA and protein components undergo highly ordered, tightly coordinated rearrangements in order to carry out intron recognition and splicing catalysis. Studies using the genetically tractable unicellular eukaryote budding yeast (Saccharomyces cerevisiae) have played an instrumental role in deciphering splicing mechanisms. In this chapter, we discuss how yeast genetics has been used to deepen our understanding of the mechanism of splicing and explore the potential for future mechanistic insights using S. cerevisiae as an experimental tool.
前体信使核糖核酸（pre-mRNA）剪接是真核基因表达中的关键步骤，它涉及从前体信使核糖核酸中去除非编码内含子序列，并将剩余的外显子序列连接起来以形成成熟的信使核糖核酸。剪接由一种称为剪接体的大型核糖核蛋白复合体执行。自20世纪70年代首次描述前体信使核糖核酸剪接反应以来，精细的遗传学和生物化学研究表明，催化该反应的酶——剪接体，是一种极其动态的大分子机器，其RNA和蛋白质组分经历高度有序、紧密协调的重排，以便进行内含子识别和剪接催化。利用遗传上易于操作的单细胞真核生物酿酒酵母进行的研究，在解读剪接机制方面发挥了重要作用。在本章中，我们将讨论如何利用酵母遗传学来加深我们对剪接机制的理解，并探讨以酿酒酵母作为实验工具获取未来机制性见解的潜力。
