Madjar J J, Nielsen-Smith K, Frahm M, Roufa D J
Proc Natl Acad Sci U S A. 1982 Feb;79(4):1003-7. doi: 10.1073/pnas.79.4.1003.
Chinese hamster ovary cell mutants resistant to the translational inhibitor emetine (Emt B mutants) express a single genetically altered 40S ribosomal subunit protein, S14. that appears to account for their drug resistance. To determine whether Emt B mutation affect the structural gene for the S14 protein, we isolated mRNAs from several wild-type and Emt B mutant clones. We translated the mRNAs in a cell-free system derived from wheat germ, and we discerned the biosynthesis of 59 ribosomal proteins, including S14. In every case, poly(A)+ mRNA programmed the cell-free system to synthesize an S14 protein electrophoretically identical to the S14 extracted from the ribosomes of the corresponding cell line. Messages from two Emt B mutants (Emr-2 and Emr-2-2) specified S14s that were electrophoretically distinct from the wild-type protein. Thus, Emt B mutations were expressed in mutant cell mRNAs, apparently reflecting mutagen-induced changes in S14 structural genes.
对翻译抑制剂吐根碱具有抗性的中国仓鼠卵巢细胞突变体(Emt B突变体)表达一种单一的、基因发生改变的40S核糖体亚基蛋白S14,这似乎是它们产生耐药性的原因。为了确定Emt B突变是否影响S14蛋白的结构基因,我们从几个野生型和Emt B突变体克隆中分离出mRNA。我们在源自小麦胚芽的无细胞系统中翻译这些mRNA,并识别出59种核糖体蛋白的生物合成,包括S14。在每种情况下,聚腺苷酸加尾的mRNA使无细胞系统合成出电泳图谱与从相应细胞系核糖体中提取的S14相同的S14蛋白。来自两个Emt B突变体(Emr-2和Emr-2-2)的信使RNA所编码的S14在电泳图谱上与野生型蛋白不同。因此,Emt B突变在突变体细胞的mRNA中得以表达,这显然反映了诱变导致的S14结构基因的变化。