Stansfield I, Tuite M F
Research School of Biosciences, University of Kent, Canterbury, UK.
Curr Genet. 1994 May;25(5):385-95. doi: 10.1007/BF00351776.
The study of translational termination in yeast has been approached largely through the identification of a range of mutations which either increase or decrease the efficiency of stop-codon recognition. Subsequent cloning of the genes encoding these factors has identified a number of proteins important for maintaining the fidelity of termination, including at least three ribosomal proteins (S5, S13, S28). Other non-ribosomal proteins have been identified by mutations which produce gross termination-accuracy defects, namely the SUP35 and SUP45 gene products which have closely-related higher eukaryote homologues (GST1-h and SUP45-h respectively) and which can complement the corresponding defective yeast proteins, implying that the yeast ribosome may be a good model for the termination apparatus existing in higher translation systems. While the yeast mitochondrial release factor has been cloned (Pel et al. 1992), the corresponding cytosolic RF has not yet been identified. It seems likely, however, that the identification of the gene encoding eRF could be achieved using a multicopy antisuppressor screen such as that employed to clone the E. coli prfA gene (Weiss et al. 1984). Identification of the yeast eRF and an investigation of its interaction with other components of the yeast translational machinery will no doubt further the definition of the translational termination process. While a large number of mutations have been isolated in which the efficiency of termination-codon recognition is impaired, it seems probable that a proportion of mutations within this class will comprise those where the accuracy of 'A' site codon-anticodon interaction is compromised: such defects would also have an effect on termination-codon suppression, allowing mis- or non-cognate tRNAs to bind stop-codons, causing nonsense suppression. The remainder of mutations affecting termination fidelity should represent mutations in genes coding for components of the termination apparatus, including the eRF: these mutations reduce the efficiency of termination, allowing nonsense suppression by low-efficiency natural suppressor tRNAs. Elucidation of the mechanism of termination in yeast will require discrimination between these two classes of mutations, thus allowing definition of termination-specific gene products.
对酵母中转录终止的研究主要通过鉴定一系列增加或降低终止密码子识别效率的突变来进行。随后对编码这些因子的基因进行克隆,确定了一些对维持终止保真度很重要的蛋白质,包括至少三种核糖体蛋白(S5、S13、S28)。其他非核糖体蛋白已通过产生严重终止准确性缺陷的突变鉴定出来,即SUP35和SUP45基因产物,它们分别具有与高等真核生物密切相关的同源物(分别为GST1-h和SUP45-h),并且可以补充相应的有缺陷的酵母蛋白,这意味着酵母核糖体可能是高等翻译系统中存在的终止装置的良好模型。虽然酵母线粒体释放因子已被克隆(佩尔等人,1992年),但相应的胞质RF尚未确定。然而,似乎可以使用多拷贝反抑制筛选来实现对编码eRF的基因的鉴定,例如用于克隆大肠杆菌prfA基因的筛选方法(魏斯等人,1984年)。鉴定酵母eRF并研究其与酵母翻译机制其他成分的相互作用无疑将进一步明确转录终止过程。虽然已经分离出大量终止密码子识别效率受损的突变,但这类突变中可能有一部分包括“A”位点密码子-反密码子相互作用准确性受损的突变:此类缺陷也会对终止密码子抑制产生影响,使错配或非同源tRNA结合终止密码子,导致无义抑制。影响终止保真度的其余突变应代表编码终止装置成分(包括eRF)的基因突变:这些突变会降低终止效率,使低效的天然抑制tRNA产生无义抑制。阐明酵母中的终止机制需要区分这两类突变,从而确定终止特异性基因产物。