Fraser James A, Davis Meryl A, Hynes Michael J
Department of Genetics, University of Melbourne, Victoria 3010, Australia.
Appl Environ Microbiol. 2002 Jun;68(6):2802-8. doi: 10.1128/AEM.68.6.2802-2808.2002.
Aspergillus nidulans is a saprophytic ascomycete that utilizes a wide variety of nitrogen sources. We identified a sequence from A. nidulans similar to the glutathione S-transferase-like nitrogen regulatory domain of Saccharomyces cerevisiae Ure2. Cloning and sequencing of the gene, designated gstA, revealed it to be more similar to URE2 than the S. cerevisiae glutathione S-transferases. However, creation and analysis of a gstA deletion mutant revealed that the gene does not participate in nitrogen metabolite repression. Instead, it encodes a functional theta class glutathione S-transferase that is involved in resistance to a variety of xenobiotics and metals and confers susceptibility to the systemic fungicide carboxin. Northern analysis showed that gstA transcription is strongly activated upon exposure to 1-chloro-2,4-dinitrobenzene and weakly activated by oxidative stress or growth on galactose as a carbon source. These results suggest that nitrogen metabolite repression in A. nidulans does not involve a homolog of the S. cerevisiae URE2 gene and that the global nitrogen regulatory system differs significantly in these two fungi.
构巢曲霉是一种腐生性子囊菌,可利用多种氮源。我们从构巢曲霉中鉴定出一个与酿酒酵母Ure2的谷胱甘肽S - 转移酶样氮调节结构域相似的序列。对该基因(命名为gstA)进行克隆和测序后发现,它与URE2的相似性高于酿酒酵母的谷胱甘肽S - 转移酶。然而,对gstA缺失突变体的构建和分析表明,该基因不参与氮代谢物阻遏。相反,它编码一种功能性的θ类谷胱甘肽S - 转移酶,参与对多种异生物质和金属的抗性,并赋予对系统性杀菌剂羧菌灵的敏感性。Northern分析表明,暴露于1 - 氯 - 2,4 - 二硝基苯时,gstA转录被强烈激活,而氧化应激或在以半乳糖作为碳源生长时则被弱激活。这些结果表明,构巢曲霉中的氮代谢物阻遏不涉及酿酒酵母URE2基因的同源物,并且这两种真菌的全局氮调节系统存在显著差异。