Transcription factor CCG-8 plays a pivotal role in azole adaptive responses of Neurospora crassa by regulating intracellular azole accumulation
Transcription factor CCG-8 plays a pivotal role in azole adaptive responses of Neurospora crassa by regulating intracellular azole accumulation
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转录因子CCG-8通过调节细胞内唑类积累在粗糙脉孢菌的唑类适应性反应中发挥关键作用
DOI：
10.1007/s00294-018-0924-7
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发表时间：
2019-01
期刊：
影响因子：
2.5
通讯作者：
Sun Xianyun
中科院分区：
文献类型：
--
作者：
Xue Wei;Yin Yajing;Ismail Fatima;Hu Chengcheng;Zhou Mi;Cao Xianhe;Li Shaojie;Sun Xianyun
唑类是临床和农业中应用最广泛的控制真菌感染的抗真菌药物。真菌可以通过快速激活一些基因的转录来适应唑类胁迫，其中一些基因可以提高对唑类药物的抗性。我们已经报道了转录因子CCG-8作为一种新的调节因子，在粗枝脉孢菌和黄萎病镰刀菌中作为一种新的调节因子来适应抗菌唑胁迫。在这项研究中，我们以粗毛拟青霉为模型，进一步研究CCG-8促进真菌对唑类胁迫适应的机制。而CCG-8基因的缺失使粗枝杆菌对唑类药物高度敏感，而CCG-8高表达菌株对唑类药物的抗性强于野生型，进一步证实了CCG-8基因在抗真菌唑类药物适应中的积极作用。液-质联用分析表明，与野生型相比，CCG-8基因缺失导致麦角甾醇生物合成减少，细胞内有毒甾醇14α-甲基-3，6-二醇和酮康唑(KTC)的积累增加，而酮康唑的细胞内积累减少。为了分析CCG-8对唑类外排的影响，我们测试了预测的多药转运体对唑类耐药性的贡献，发现CDR4是粗糙奈瑟菌中唑类外排的主要贡献者。有趣的是，在CCG-8缺失突变体中过表达cdr4或erg11可以恢复其超敏表型，并且过表达cdr4可以降低细胞内KTC的水平。而CCG-8和cdr4双突变体对药物摄取的敏感性高于单个突变体，提示在CCG-8缺失突变体中，药物外排泵CDR4对细胞内氮唑蓄积的贡献较小，CCG-8可能调节药物摄取。综上所述，我们的结果表明，CCG-8通过调节药物在细胞内的积累，在粗糙奈瑟菌的唑类适应性反应中发挥了关键作用。
Azoles are the most widely used antifungals for controlling fungal infections in clinic and agriculture. Fungi can adapt to azole stress by rapidly activating the transcription of a number of genes, and some of these genes can elevate resistance to azoles. We had reported the transcription factor CCG-8 as a new regulator in the adaptation to antifungal azole stress in Neurospora crassa and Fusarium verticillioides. In this study, we further investigate the mechanisms by which CCG-8 promotes fungal adaptation to azole stress using N. crassa as a model. While deletion of ccg-8 made N. crassa hypersensitive to azoles, ccg-8 overexpression strain was more resistant to azoles than wild type, which further confirmed the positive role of ccg-8 in the adaptation to antifungal azoles. Liquid chromatography-mass spectrometry analysis showed that deletion of ccg-8 resulted in decrease of ergosterol biosynthesis, and high accumulation of toxic sterol 14α-methyl-3,6-diol and ketoconazole (KTC) in the cells, whereas intracellular accumulation of ketoconazole was decreased in the ccg-8 overexpression strain as compared to wild type. For analyzing the effect of CCG-8 on azole export, we tested the contribution of predicted multidrug transporters to azole resistance and found that CDR4 is the major contributor for azole efflux in N. crassa. Interestingly, overexpression of cdr4 or erg11 in the ccg-8 deletion mutant restored its hypersensitive phenotype and overexpression of cdr4 can reduce the level of intracellular KTC. However, the double mutant of ccg-8 and cdr4 was more sensitive than each single mutant, suggesting that drug efflux pump CDR4 plays less contribution for intracellular azole accumulation in the ccg-8 deletion mutant, and CCG-8 may regulate drug uptake. Together, our results revealed that CCG-8 plays a pivotal role in azole adaptive responses of N. crassa by regulating the drug accumulation in the cells.
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影响因子：
6.7
作者：
Mansfield BE;Oltean HN;Oliver BG;Hoot SJ;Leyde SE;Hedstrom L;White TC
通讯作者：
White TC
影响因子：
6.7
作者：
Hagiwara D;Miura D;Shimizu K;Paul S;Ohba A;Gonoi T;Watanabe A;Kamei K;Shintani T;Moye-Rowley WS;Kawamoto S;Gomi K
通讯作者：
Gomi K
影响因子：
2.5
作者：
Yu Zhang;Zhen-ying Zhang;Xinyu Zhang;Hanxing Zhang;Xianyun Sun;Chengcheng Hu;Shaojie Li
通讯作者：
Yu Zhang;Zhen-ying Zhang;Xinyu Zhang;Hanxing Zhang;Xianyun Sun;Chengcheng Hu;Shaojie Li
影响因子：
4.8
作者：
Kolaczkowski, M;vanderRest, M;Goffeau, A
通讯作者：
Goffeau, A
DOI：
--
发表时间：
1991-01
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
M. White;J. Hirsch;S. Henry
通讯作者：
M. White;J. Hirsch;S. Henry