Finnegan P M, Payne M J, Keramidaris E, Lukins H B
Department of Biochemistry, Monash University, Clayton, Victoria, Australia.
Curr Genet. 1991 Jul;20(1-2):53-61. doi: 10.1007/BF00312765.
The temperature-conditional pet mutant, ts379, of Saccharomyces cerevisiae fails to synthesize mitochondrial ATP synthase subunit 9 at the restrictive temperature due to mutation of a single nuclear locus, AEP2. The inability to synthesize subunit 9 correlates with a lowered accumulation of the cognate oli1 mRNA indicating that the AEP2 product is involved in oli1 transcript maturation or stabilization. The AEP2 gene has been isolated in this study from a wild-type yeast genomic library by genetic complementation of ts379 at the restrictive temperature. A 1,740 nucleotide open-reading frame was observed that encodes a basic, hydrophilic protein of 67,534 Da which possesses a putative mitochondrial address signal. Disruption of chromosomal DNA within this reading frame produced a non-conditional respiratory mutant unable to synthesize subunit 9, identifying the AEP2 gene. Hybridization analyses indicate that AEP2 is located on chromosome XIII and produces a 2.1 kb poly(A)+ transcript. Two additional open-reading frames were found in close proximity to that of AEP2. The three open-reading frames shared no significant homology with entries in several data bases.
酿酒酵母的温度条件性pet突变体ts379,由于单一位点AEP2发生突变,在限制温度下无法合成线粒体ATP合酶亚基9。无法合成亚基9与同源oli1 mRNA积累减少相关,这表明AEP2产物参与oli1转录本的成熟或稳定。在本研究中,通过在限制温度下对ts379进行遗传互补,从野生型酵母基因组文库中分离出了AEP2基因。观察到一个1740个核苷酸的开放阅读框,其编码一个67534 Da的碱性亲水性蛋白,该蛋白具有一个假定的线粒体定位信号。破坏该阅读框内的染色体DNA产生了一个非条件性呼吸突变体,该突变体无法合成亚基9,从而确定了AEP2基因。杂交分析表明,AEP2位于第十三号染色体上,并产生一个2.1 kb的聚腺苷酸化转录本。在AEP2附近还发现了另外两个开放阅读框。这三个开放阅读框与几个数据库中的条目没有明显的同源性。