YEAST ADENYLATE KINASE IS ACTIVE SIMULTANEOUSLY IN MITOCHONDRIA AND CYTOPLASM AND IS REQUIRED FOR NON-FERMENTATIVE GROWTH
YEAST ADENYLATE KINASE IS ACTIVE SIMULTANEOUSLY IN MITOCHONDRIA AND CYTOPLASM AND IS REQUIRED FOR NON-FERMENTATIVE GROWTH
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DOI：
10.1111/j.1432-1033.1988.tb14469.x
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发表时间：
1988-12-15
期刊：
影响因子：
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通讯作者：
MAGDOLEN, V
中科院分区：
文献类型：
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作者：
BANDLOW, W;STROBEL, G;MAGDOLEN, V
在单倍体细胞中，酵母腺苷酸激酶的单拷贝结构基因（长版本）被一个中断的无功能等位基因所取代是可以耐受的。由于腺苷酸激酶活性是细胞活力的先决条件，单倍体破坏突变体的存活表明酵母菌中存在腺苷酸激酶同工酶，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，表明互补仅在发酵条件下发生。即使在葡萄糖中，突变体的生长也是缓慢的。腺苷酸激酶活性主要存在于野生型酵母的线粒体和细胞质中。这种破坏完全破坏了线粒体的活性，而细胞质部分则保留了约10%。一种针对酵母线粒体腺苷酸激酶的抗体可以识别野生型线粒体和细胞质中的交叉反应物质，但在每个各自的突变部分中都不能这样做。结果表明，酵母腺苷酸激酶（长链AKY2）在野生型的线粒体和细胞质中同时存在并具有活性。然而，它缺乏进入线粒体的可切割的前序列。第二种较小的同工酶，由一个单独的基因编码，只存在于细胞质中。
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 bot in mitochondira 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 of import into mitochondria. A second, minor isozyme, encoded by a separate gene, is present exclusively in the cytoplasm.