Gliotoxin effects on fungal growth: Mechanisms and exploitation
Gliotoxin effects on fungal growth: Mechanisms and exploitation
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DOI：
10.1016/j.fgb.2012.02.003
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发表时间：
2012-04-01
影响因子：
3
通讯作者：
Doyle, Sean
中科院分区：
文献类型：
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作者：
Carberry, Stephen;Molloy, Emer;Doyle, Sean
尽管最初对其抗真菌特性进行了研究，但实际上人们对胶霉毒素对烟曲霉和其他真菌的影响知之甚少。我们观察到，在胶霉毒素限制的生长条件下，烟曲霉暴露于外源胶霉毒素（14 μg/ml），会导致 27 种蛋白质表达显着改变（上调和下调 >1.9 倍；p < 0.05），包括 Cu、Zn 超氧化物歧化酶的从头表达、过敏原 Asp f3 表达上调以及过氧化氢酶和过氧化物酶水平下调。在缺乏胶霉毒素氧化还原酶（一种胶霉毒素自我保护基因）的烟曲霉 Delta gliT 中，谷胱甘肽 GSH 水平显着升高（p < 0.05），并伴有对二酰胺的抗性。酿酒酵母缺失株（Delta sod1 和 Delta yap1）对外源胶霉毒素过敏，而 Delta gsh1 具有抗性。还观察到胶霉毒素介导的（5μg/ml）对构巢曲霉、土曲霉、黑曲霉、异旋旋孢菌和粗糙脉孢菌的显着生长抑制（p<0.001）。黄曲霉、禾谷镰刀菌和米曲霉的生长在胶霉毒素（10 μg/ml）下显着受到抑制（p < 0.001），表明真菌之间对胶霉毒素的敏感性不同。将 gliT 重新引入烟曲霉 Delta gliT 的不同位点（ctsD；AFUA_4G07040，一种天冬氨酸蛋白酶），并选择胶霉毒素，无需使用额外的抗生素选择标记即可促进 ctsD 的删除。 ctsD 表达的缺失伴随着 gliT 表达的恢复以及对胶霉毒素的抗性。因此，我们建议 gliT/gliotoxin 作为真菌转化的有用选择标记系统。最后，我们建议将胶霉毒素敏感性测定纳入所有未来的真菌功能基因组研究中。 (C) 2012 Elsevier Inc. 保留所有权利。
Although initially investigated for its antifungal properties, little is actually known about the effect of gliotoxin on Aspergillus fumigatus and other fungi. We have observed that exposure of A. fumigatus to exogenous gliotoxin (14 mu g/ml), under gliotoxin-limited growth conditions, results in significant alteration of the expression of 27 proteins (up- and down-regulated >1.9-fold; p < 0.05) including de novo expression of Cu, Zn superoxide dismutase, up-regulated allergen Asp f3 expression and down-regulated catalase and a peroxiredoxin levels. Significantly elevated glutathione GSH levels (p < 0.05), along with concomitant resistance to diamide, were evident in A. fumigatus Delta gliT, lacking gliotoxin oxidoreductase, a gliotoxin self-protection gene. Saccharomyces cerevisiae deletents (Delta sod1 and Delta yap1) were hypersensitive to exogenous gliotoxin, while Delta gsh1 was resistant. Significant gliotoxin-mediated (5 mu g/ml) growth inhibition (p < 0.001) of Aspergillus nidulans, Aspergillus terreus, Aspergillus niger, Cochliobolus heterostrophus and Neurospora crassa was also observed. Growth of Aspergillus flavus, Fusarium graminearum and Aspergillus oryzae was significantly inhibited (p < 0.001) at gliotoxin (10 mu g/ml), indicating differential gliotoxin sensitivity amongst fungi. Re-introduction of gliT into A. fumigatus Delta gliT, at a different locus (ctsD; AFUA_4G07040, an aspartic protease), with selection on gliotoxin, facilitated deletion of ctsD without use of additional antibiotic selection markers. Absence of ctsD expression was accompanied by restoration of gliT expression, and resistance to gliotoxin. Thus, we propose gliT/gliotoxin as a useful selection marker system for fungal transformation. Finally, we suggest incorporation of gliotoxin sensitivity assays into all future fungal functional genomic studies. (C) 2012 Elsevier Inc. All rights reserved.