Fried H M, Warner J R
Proc Natl Acad Sci U S A. 1981 Jan;78(1):238-42. doi: 10.1073/pnas.78.1.238.
Yeast cells sensitive to the eukaryotic protein synthesis inhibitor trichodermin have been transformed with autonomously replicating recombinant plasmids carrying DNA fragments of the genome of a trichodermin-resistant yeast strain. After selection for trichodermin-resistant cells, several transformants yielded a plasmid containing a 13.5-kilobase (kb) DNA fragment that encodes the trichodermin resistance gene, tcm1, and the gene for ribosomal protein L3, the largest of the yeast ribosomal proteins. Cells carrying this plasmid are resistant to trichodermin and to the related drug verrucarin A as well as to the unrelated drug anisomycin. This pattern of resistance is similar to that exhibited by strains carrying a chromosomal copy of tcm1. Moreover, polyribosomes prepared from transformed cells are resistant to trichodermin when tested in an in vitro protein synthesis assay. Subcloning of the 13.5-kb DNA fragment revealed that the gene for tcm1 and the gene for protein L3 are contained within a 3.2-kb segment. These results suggest that the gene for trichodermin resistance in yeast specifies ribosomal protein L3.
对真核生物蛋白质合成抑制剂木霉菌素敏感的酵母细胞，已用携带抗木霉菌素酵母菌株基因组DNA片段的自主复制重组质粒进行了转化。在筛选出抗木霉菌素的细胞后，几个转化子产生了一种质粒，该质粒含有一个13.5千碱基（kb）的DNA片段，该片段编码木霉菌素抗性基因tcm1和核糖体蛋白L3的基因，L3是酵母核糖体蛋白中最大的一种。携带这种质粒的细胞对木霉菌素、相关药物疣孢菌素A以及无关药物茴香霉素均具有抗性。这种抗性模式与携带tcm1染色体拷贝的菌株所表现出的抗性模式相似。此外，在体外蛋白质合成试验中测试时，从转化细胞制备的多核糖体对木霉菌素具有抗性。对13.5-kb DNA片段进行亚克隆后发现，tcm1基因和蛋白L3基因位于一个3.2-kb的片段内。这些结果表明，酵母中木霉菌素抗性基因指定了核糖体蛋白L3。
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