Fox T D, Staempfli S
Proc Natl Acad Sci U S A. 1982 Mar;79(5):1583-7. doi: 10.1073/pnas.79.5.1583.
A polypeptide chain-terminating mutation in the yeast mitochondrial oxi 1 gene has been shown to be an ochre (TAA) mutation by DNA sequence analysis. Mitochondrially inherited revertants of this mutation include two types: In the first, the ochre codon has been changed to a sense codon by further mutation in the oxi 1 gene while, in the second, the ochre codon is still present, indicating the occurrence of an extrageneic ochre suppressor mutation. This mitochondrial ochre suppressor, termed MSU1, has been "cloned" in rho- strains of yeast and tested against other oxi 1 mutations. Several additional mutations are also suppressible, and those examined so far are also ochre mutations. MSU1 does not suppress known frameshift or missense mutations at oxi 1. Isoelectric focusing of the gene product (cytochrome oxidase subunit II) from a suppressed-mutant strain indicates that suppression does not involve insertion of charged amino acids. Physical mapping of the mtDNA retained in the MSU1-carrying rho- clones localizes the suppressor mutation to the gene coding the 15S rRNA or a site not more than 300 base pairs from it. No known tRNA genes occur this close to the 15S rRNA gene, and mtDNA from a suppressor-carrying rho- does not hybridize detectably to mitochondrial tRNAs. These results suggest that MSU1 may be an alteration in the 15S rRNA.
通过DNA序列分析表明,酵母线粒体oxi 1基因中的一个多肽链终止突变是一个赭石型(TAA)突变。该突变的线粒体遗传回复突变体包括两种类型:第一种,oxi 1基因中的进一步突变将赭石密码子改变为有义密码子;而第二种,赭石密码子仍然存在,表明发生了基因外赭石抑制突变。这种线粒体赭石抑制子,称为MSU1,已在酵母的rho-菌株中“克隆”,并针对其他oxi 1突变进行了测试。其他几个突变也可被抑制,并且到目前为止所检测的那些突变也是赭石突变。MSU1不能抑制oxi 1处已知的移码或错义突变。对来自一个被抑制突变菌株的基因产物(细胞色素氧化酶亚基II)进行等电聚焦表明,抑制作用不涉及带电荷氨基酸的插入。保留在携带MSU1的rho-克隆中的mtDNA的物理图谱将抑制突变定位到编码15S rRNA的基因或距其不超过300个碱基对的位点。在距15S rRNA基因如此近的位置没有已知的tRNA基因,并且来自携带抑制子的rho-的mtDNA不能与线粒体tRNA发生可检测到的杂交。这些结果表明,MSU1可能是15S rRNA中的一种改变。