Bandlow W, Strobel G, Zoglowek C, Oechsner U, Magdolen V
Institute for Genetics and Microbiology, University of Munich, Federal Republic of Germany.
Eur J Biochem. 1988 Dec 15;178(2):451-7. doi: 10.1111/j.1432-1033.1988.tb14469.x.
Displacement of the single copy structural gene for yeast adenylate kinase (long version) by a disrupted nonfunctional allele is tolerated in haploid cells. Since adenylate kinase activity is a pre-requisite for cell viability, the survival of haploid disruption mutants is indicative of the presence of an adenylate kinase isozyme in yeast, capable of forming ADP from AMP and, thus, of complementing the disrupted allele. The phenotype of these disruption mutants is pet, showing that complementation occurs only under fermentative conditions. Even on glucose, growth of the disruption mutants is slow. Adenylate kinase activity is found both in mitochondria and cytoplasm of wild type yeast. The disruption completely destroys the activity in mitochondria, whereas in the cytoplasmic fraction about 10% is retained. An antibody raised against yeast mitochondrial adenylate kinase recognizes cross-reacting material both in mitochondria and cytoplasm of the wild type, but fails to do so in each of the respective mutant fractions. The data indicate that yeast adenylate kinase (long version, AKY2) simultaneously occurs and is active in mitochondria and cytoplasm of the wild type. Nevertheless, it lacks a cleavable pre-sequence for import into mitochondria. A second, minor isozyme, encoded by a separate gene, is present exclusively in the cytoplasm.
单拷贝的酵母腺苷酸激酶结构基因(长版本)被一个失活的无功能等位基因取代后,在单倍体细胞中是可以耐受的。由于腺苷酸激酶活性是细胞存活的先决条件,单倍体破坏突变体的存活表明酵母中存在一种腺苷酸激酶同工酶,它能够从AMP形成ADP,从而补充被破坏的等位基因。这些破坏突变体的表型是“pet”,表明互补作用仅在发酵条件下发生。即使在葡萄糖上,破坏突变体的生长也很缓慢。腺苷酸激酶活性在野生型酵母的线粒体和细胞质中均有发现。这种破坏完全消除了线粒体中的活性,而在细胞质部分中仍保留约10%。针对酵母线粒体腺苷酸激酶产生的抗体能识别野生型线粒体和细胞质中的交叉反应物质,但在各自的突变体部分中均未识别到。数据表明,酵母腺苷酸激酶(长版本,AKY2)在野生型的线粒体和细胞质中同时存在且具有活性。然而,它缺乏可裂解的导入线粒体的前导序列。由另一个单独基因编码的第二种次要同工酶仅存在于细胞质中。