VERSATILE VECTORS TO STUDY RECODING - CONSERVATION OF RULES BETWEEN YEAST AND MAMMALIAN-CELLS
VERSATILE VECTORS TO STUDY RECODING - CONSERVATION OF RULES BETWEEN YEAST AND MAMMALIAN-CELLS
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DOI：
10.1093/nar/23.9.1557
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发表时间：
1995-05-11
影响因子：
14.9
通讯作者：
CASSAN, M
中科院分区：
文献类型：
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作者：
STAHL, G;BIDOU, L;CASSAN, M
在许多病毒和转座子中，某些基因的表达需要遗传密码的替代读取，也称为重新编码。此类事件取决于特定的 mRNA 序列，并可能导致读框内终止密码子或 +1 或 -1 移码。在这里，我们通过建立可用于研究两个物种重新编码的多功能载体，解决了酿酒酵母和哺乳动物细胞之间重新编码规则的保守问题。我们首先通过分析 Ty1 转座子的 +1 移码位点来评估该载体。然后在酵母和哺乳动物细胞中测试了来自高等生物体的两个序列：人类免疫缺陷病毒1型（HIV-1）的gag-pol连接（-1移码位点）和烟草花叶病毒复制酶顺反子的终止密码子区域（UAG通读位点）。在缺乏 HIV-1 茎环刺激信号的情况下观察到移码减少。总之，这些数据表明一些重编码事件的机制在低等真核生物和高等真核生物之间是保守的，从而允许使用酿酒酵母作为模型系统来研究来自高等生物的靶序列的重编码。
In many viruses and transposons, expression of some genes requires alternative reading of the genetic code, also called recoding, Such events depend on specific mRNA sequences and can lead to read through of an in-frame stop codon or to +1 or -1 frameshifting. Here, we addressed the issue of conservation of recoding rules between the yeast Saccharomyces cerevisiae and mammalian cells by establishing a versatile vector that can be used to study recoding in both species, We first assessed this vector by analysing the site of +1 frameshift of the Ty1 transposon. Two sequences from higher organisms were then tested in both yeast and mammalian cells: the gag-pol junction of human immunodeficiency virus type 1 (HIV-1) (a site of -1 frameshift), and the stop codon region of the replicase cistron from the tobacco mosaic virus (a site of UAG read through), We show that both sequences direct a high level of recoding in yeast, Furthermore, different mutations of the target sequences have similar effects on recoding in yeast and in mouse cells, Most notably, a strong decrease of frameshifting was observed in the absence of the HIV-1 stem-loop stimulatory signal, Taken together, these data suggest that mechanisms of some recoding events are conserved between lower and higher eukaryotes, thus allowing the use of S.cerevisiae as a model system to study recoding on target sequences from higher organisms.