Fried H M, Pearson N J, Kim C H, Warner J R
J Biol Chem. 1981 Oct 10;256(19):10176-83.
We have isolated recombinant lambda phage carrying the genes for 14 of the ribosomal proteins of the yeast Saccharomyces cerevisiae. Analysis of these and of the plasmid carrying the gene tcm1, which codes for the ribosomal protein responsible for resistance to trichodermin, demonstrates that in general the genes for ribosomal proteins are unlinked. One exceptional recombinant carries the genes for two ribosomal proteins within a 2-kilobase region. DNA fragments bearing individual ribosomal protein genes were used to probe restriction digests of the yeast genome to determine whether any of the genes were duplicated. Only 3 of 12 of the genes are present unequivocally as a single copy. Similar fragments were used to probe blots of mRNA separated on denaturing agarose gels to determine the size of the mRNA for each protein. In each case, the mRNA is near the minimum size necessary to code for its protein. In certain temperature-sensitive mutants which fail to synthesize functional mRNA for ribosomal protein, Rosbash et al. (Rosbash, M., Harris, P. K. W., Woolford, J., and Teem, J. L. (1981) Cell, 24, 679-686) have demonstrated the accumulation of a larger RNA molecule, homologous to a ribosomal protein gene, that appears to be a transcript which retains an intervening sequence. We find that for 8 of the 11 ribosomal protein genes examined, a larger molecule accumulates in such a mutant strain, suggesting that in general transcripts of ribosomal protein genes may have introns.
我们已经分离出携带酿酒酵母14种核糖体蛋白基因的重组λ噬菌体。对这些基因以及携带tcm1基因的质粒(该基因编码对木霉菌素具有抗性的核糖体蛋白)的分析表明,核糖体蛋白基因通常是不连锁的。一个特殊的重组体在一个2千碱基区域内携带了两个核糖体蛋白的基因。携带单个核糖体蛋白基因的DNA片段被用于探测酵母基因组的限制性酶切片段,以确定是否有任何基因是重复的。12个基因中只有3个明确以单拷贝形式存在。类似的片段被用于探测在变性琼脂糖凝胶上分离的mRNA印迹,以确定每种蛋白质的mRNA大小。在每种情况下,mRNA接近编码其蛋白质所需的最小大小。在某些不能合成核糖体蛋白功能性mRNA的温度敏感突变体中,罗斯巴什等人(罗斯巴什,M.,哈里斯,P.K.W.,伍尔福德,J.,和蒂姆,J.L.(1981年)《细胞》,24卷,679 - 686页)已经证明积累了一种与核糖体蛋白基因同源的更大的RNA分子,它似乎是一个保留了间隔序列的转录本。我们发现,在所检测的11个核糖体蛋白基因中的8个基因,在这样的突变菌株中会积累更大的分子,这表明核糖体蛋白基因的转录本通常可能含有内含子。