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首页> 外文期刊>G3: Genes, Genomes, Genetics >Identification of Genes Required for Alternative Oxidase Production in the Neurospora crassa Gene Knockout Library
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Identification of Genes Required for Alternative Oxidase Production in the Neurospora crassa Gene Knockout Library

机译：鉴定神经孢霉基因敲除文库中替代氧化酶生产所需的基因

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pThe alternative oxidase (AOX) of iNeurospora crassa/i transfers electrons from ubiquinol to oxygen. The enzyme is not expressed under normal conditions. However, when the function of the standard electron transport chain is compromised, AOX is induced, providing cells with a means to continue respiration and growth. Induction of the enzyme represents a form of retrograde regulation because AOX is encoded by a nuclear gene that responds to signals produced from inefficiently functioning mitochondria. To identify genes required for AOX expression, we have screened the iN. crassa/i gene knockout library for strains that are unable to grow in the presence of antimycin A, an inhibitor of complex III of the standard electron transport chain. From the 7800 strains containing knockouts of different genes, we identified 62 strains that have reduced levels of AOX when grown under conditions known to induce the enzyme. Some strains have virtually no AOX, whereas others have only a slight reduction of the protein. A broad range of seemingly unrelated functions are represented in the knockouts. For example, we identified transcription factors, kinases, the mitochondrial import receptor Tom70, three subunits of the COP9 signalosome, a monothiol glutaredoxin, and several hypothetical proteins as being required for wild-type levels of AOX production. Our results suggest that defects in many signaling or metabolic pathways have a negative effect on AOX expression and imply that complex systems control production of the enzyme./p
机译：> Creourospora crassa 的替代氧化酶（AOX）将电子从泛醇转移到氧气。该酶在正常条件下不表达。但是，当标准电子传输链的功能受损时，会诱导AOX，为细胞提供继续呼吸和生长的方式。酶的诱导代表逆行调节的一种形式，因为AOX是由核基因编码的，该核基因对功能低下的线粒体产生的信号作出反应。为了鉴定AOX表达所需的基因，我们筛选了N。 crassa 基因敲除文库，用于在抗霉素A（标准电子传输链复合物III的抑制剂）存在下无法生长的菌株。从包含不同基因敲除的7800株菌株中，我们鉴定出62株在已知诱导酶的条件下生长时AOX含量降低的菌株。有些菌株实际上没有AOX，而另一些菌株的蛋白质仅略有减少。剔除中包含了许多看似无关的功能。例如，我们确定了野生型AOX产生所需的转录因子，激酶，线粒体导入受体Tom70，COP9信号体的三个亚基，一硫醇戊二醛毒素和几种假设的蛋白质。我们的结果表明，许多信号或代谢途径中的缺陷对AOX的表达都具有负面影响，这暗示着复杂的系统会控制酶的产生。

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