Illumina identification of RsrA, a conserved C2H2 transcription factor coordinating the NapA mediated oxidative stress signaling pathway in Aspergillus.
Illumina identification of RsrA, a conserved C2H2 transcription factor coordinating the NapA mediated oxidative stress signaling pathway in Aspergillus.
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DOI：
10.1186/1471-2164-15-1011
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发表时间：
2014-11-22
期刊：
影响因子：
4.4
通讯作者：
Keller NP
中科院分区：
文献类型：
--
作者：
Bok JW;Wiemann P;Garvey GS;Lim FY;Haas B;Wortman J;Keller NP
化学诱变筛选对鉴定参与感兴趣的生物过程的突变体很有用。然而，当使用DNA文库将突变转化为野生型的方法时，从这样的筛查中识别突变往往失败。在这里，我们分析了一个化学衍生的nidulans曲霉突变体的Illumina序列，并鉴定了一个编码C2H2转录因子的基因，称为RsrA，用于调节应激反应。RsrA在丝状真菌基因组中是保守的，当删除三种曲霉(A.nidulans、A.Huang us和A.fumigatus)的RsrA基因时，我们发现了两种保守的表型：增强了对氧化应激的抗性，减少了孢子形成过程。在所有物种中，rsrA缺失突变体对过氧化氢处理的抗性更强。对根结线虫的这一特征的深入研究表明，当暴露于过氧化氢时，RsrA的丢失导致氧化应激代谢组的几个组分的全球上调，包括NapA和ATFA的表达，这两个bZIP转录因子介导对活性氧物种(ROS)的抗性，以及硫氧还蛋白和谷胱甘肽系统中的NapA靶标。结合转录数据和ΔrsrAΔATFA和ΔrsrAΔNapA双突变体的检测，表明RsrA主要通过NapA介导的胁迫反应途径发挥作用。提出了曲霉RsrA调节ROS反应的模型。RsrA发现于曲霉基因组中的高度共线区域，协调曲霉真菌中NapA介导的氧化反应。本文的在线版本(DOI：10.1186/1471-2164-15-1011)包含补充材料，授权用户可以使用。
Chemical mutagenesis screens are useful to identify mutants involved in biological processes of interest. Identifying the mutation from such screens, however, often fails when using methodologies involving transformation of the mutant to wild type phenotype with DNA libraries. Here we analyzed Illumina sequence of a chemically derived mutant of Aspergillus nidulans and identified a gene encoding a C2H2 transcription factor termed RsrA for regulator of stress response. RsrA is conserved in filamentous fungal genomes, and upon deleting the gene in three Aspergillus species (A. nidulans, A. flavus and A. fumigatus), we found two conserved phenotypes: enhanced resistance to oxidative stress and reduction in sporulation processes. For all species, rsrA deletion mutants were more resistant to hydrogen peroxide treatment. In depth examination of this latter characteristic in A. nidulans showed that upon exposure to hydrogen peroxide, RsrA loss resulted in global up-regulation of several components of the oxidative stress metabolome including the expression of napA and atfA, the two bZIP transcription factors mediating resistance to reactive oxygen species (ROS) as well as NapA targets in thioredoxin and glutathione systems. Coupling transcriptional data with examination of ΔrsrAΔatfA and ΔrsrAΔnapA double mutants indicate that RsrA primarily operates through NapA-mediated stress response pathways. A model of RsrA regulation of ROS response in Aspergillus is presented. RsrA, found in a highly syntenic region in Aspergillus genomes, coordinates a NapA mediated oxidative response in Aspergillus fungi. The online version of this article (doi:10.1186/1471-2164-15-1011) contains supplementary material, which is available to authorized users.
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