Pierce Jacqueline B, Chafe Shawn C, Eswara Manoja B K, van der Merwe George, Mangroo Dev
Department of Molecular and Cellular Biology, University of Guelph, Guelph, Ontario, Canada.
Methods Cell Biol. 2014;122:415-36. doi: 10.1016/B978-0-12-417160-2.00019-9.
Nuclear-cytoplasmic tRNA transport involves multiple pathways that are segregated by the involvement of distinct proteins. The tRNA export process begins in the nucleolus, where the functionality of newly produced tRNAs are tested by aminoacylation, and ends with the delivery of the exported aminoacyl tRNAs to the eukaryotic elongation factor eEF-1A for utilization in protein synthesis in the cytoplasm. Recent studies have identified a number of proteins that participate in nuclear tRNA export in both yeast and mammals. However, genetic and biochemical evidence suggest that additional components, which have yet to be identified, also participate in nuclear-cytoplasmic tRNA trafficking. Here we review key strategies that have led to the identification and characterization of proteins that are involved in the nuclear tRNA export process in yeasts and mammals. The approaches described will greatly facilitate the identification and delineation of the roles of new proteins involved in nuclear export of tRNAs to the cytoplasm.
核质tRNA转运涉及多种途径,这些途径因不同蛋白质的参与而被区分开来。tRNA输出过程始于核仁,在核仁中通过氨酰化作用测试新产生tRNA的功能,最终将输出的氨酰tRNA传递给真核延伸因子eEF-1A,以供在细胞质中进行蛋白质合成时使用。最近的研究已经鉴定出许多参与酵母和哺乳动物细胞核tRNA输出的蛋白质。然而,遗传和生化证据表明,尚未被鉴定的其他成分也参与了核质tRNA的运输。在这里,我们综述了一些关键策略,这些策略促成了对酵母和哺乳动物细胞核tRNA输出过程中所涉及蛋白质的鉴定和表征。所描述的方法将极大地促进对参与tRNA从细胞核输出到细胞质的新蛋白质的作用的鉴定和描述。