Deletion of cox7c Results in Pan-Azole Resistance in Aspergillus fumigatus.
Deletion of cox7c Results in Pan-Azole Resistance in Aspergillus fumigatus.
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DOI：
10.1128/aac.00151-22
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发表时间：
2022-06-21
影响因子：
4.9
通讯作者：
中科院分区：
文献类型：
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在烟曲霉中，最普遍的唑类抗性是由唑类靶蛋白Cyp51A的突变修饰引起的，但也存在对唑类具有抗性的非cyp51A突变体，这些菌株的抗性机制仍有待探索。在这里，我们在实验室中鉴定了一种新型细胞色素 c 氧化酶 cox7c (W56*)，这是一种无义突变，并发现它会导致菌落生长减少和对多种抗真菌药物的耐药性。同时，我们发现冷藏导致分生孢子对伊曲康唑（ITC）胁迫的耐受性增加，这进一步促进了唑类抗性突变（低温保存→ITC耐受→唑类抗性）。 cox7c 的缺失或突变明确导致对麦角甾醇合成所需的抗真菌靶向酶（包括三唑、多烯和烯丙胺）的抗性，或对真菌麦角甾醇的抗性。高效液相色谱 (HPLC) 测定表明，cox7c 敲除菌株降低了细胞内伊曲康唑浓度。此外，Cox7c的缺乏导致细胞内血红素B的积累。我们验证了血红素B的内源增加或外源添加能够引发唑类抗性，这与cox7c突变体的表型抗性分析非常一致。此外，RNA测序证实了多药转运基因的转录表达水平升高。此外，cox7c 缺失菌株菌丝体中伊曲康唑诱导的活性氧生成减少表明，这种减少可能部分导致耐药性。这些发现加深了我们对烟曲霉对唑类的直接反应如何促进环境中真菌存活并解决患者或环境引起的基因突变的理解。
In Aspergillus fumigatus, the most prevalent resistance to azoles results from mutational modifications of the azole target protein Cyp51A, but there are non-cyp51A mutants resistant to azoles, and the mechanisms underlying the resistance of these strains remain to be explored. Here, we identified a novel cytochrome c oxidase, cox7c (W56*), nonsense mutation in the laboratory and found that it caused reduced colony growth and resistance to multiantifungal agents. Meanwhile, we revealed that cold storage is responsible for increased tolerance of conidia to itraconazole (ITC) stress, which further advances azole-resistant mutations (cryopreservation→ITC tolerance→azole resistance). The deletion or mutation of cox7c results explicitly in resistance to antifungal-targeting enzymes, including triazoles, polyenes, and allylamines, required for ergosterol synthesis, or resistance to fungal ergosterol. A high-performance liquid chromatography (HPLC) assay showed that the cox7c knockout strain decreased intracellular itraconazole concentration. In addition, the lack of Cox7c resulted in the accumulation of intracellular heme B. We validated that an endogenous increase in, or the exogenous addition of, heme B was capable of eliciting azole resistance, which was in good accordance with the phenotypic resistance analysis of cox7c mutants. Furthermore, RNA sequencing verified the elevated transcriptional expression levels of multidrug transport genes. Additionally, lower itraconazole-induced reactive oxygen species generation in mycelia of a cox7c-deletion strain suggested that this reduction may, in part, contribute to drug resistance. These findings increase our understanding of how A. fumigatus’s direct responses to azoles promote fungal survival in the environment and address genetic mutations that arise from patients or environments.
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影响因子：
11.8
作者：
Hagiwara D;Arai T;Takahashi H;Kusuya Y;Watanabe A;Kamei K
通讯作者：
Kamei K
影响因子：
9.8
作者：
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通讯作者：
Shoubridge, EA
DOI：
10.1016/j.bbrc.2003.09.151
发表时间：
2003-10-31
影响因子：
3.1
作者：
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通讯作者：
Rytka, J
DOI：
10.1073/pnas.2003700117
发表时间：
2020-09-08
影响因子：
11.1
作者：
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通讯作者：
Cramer, Robert A.
烟曲霉中假定的线粒体铁转运蛋白 MrsA 在唑、氧化应激反应和毒力中发挥重要作用
影响因子：
5.2
作者：
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