Accumulation of Azole Drugs in the Fungal Plant Pathogen Magnaporthe oryzae Is the Result of Facilitated Diffusion Influx.
Accumulation of Azole Drugs in the Fungal Plant Pathogen Magnaporthe oryzae Is the Result of Facilitated Diffusion Influx.
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DOI：
10.3389/fmicb.2017.01320
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发表时间：
2017
影响因子：
5.2
通讯作者：
White TC
中科院分区：
文献类型：
--
作者：
Esquivel BD;White TC
稻瘟病菌是一种农业霉菌，会导致水稻病害，造成毁灭性的作物损失。由于水稻是世界性的主要粮食作物，因此，M。对全球粮食安全构成严重威胁。杀真菌剂，包括唑类抗真菌剂，用于预防和对抗M。植物感染。唑类的靶点是CYP 51，CYP 51是一种位于内质网（ER）上的酶，是真菌麦角固醇生物合成所必需的。然而，许多基本的药物-病原体相互作用，如唑如何通过真菌细胞壁和质膜，并被运送到ER，是不清楚的。此外，在许多真菌物种中，抗真菌剂的细胞内积累减少一直被观察到是一种耐药性机制。研究药物-病原体相互作用的基础生物学可能阐明M.真菌和潜在的其他相关真菌病原体。我们描述了唑类药物在M.使用放射性标记的氟康唑摄取测定来了解唑类药物是否通过被动扩散、主动转运或易化扩散进入细胞。我们表明，唑积累不是ATP依赖性的，也不依赖于pH值依赖性的过程。相反，有证据表明唑类药物在M.通过易化扩散机制进行扩散。摄取系统对唑类或唑类化合物具有特异性，并且可以根据细胞期和生长培养基进行调节。此外，我们还发现M.用“再利用”的氯吉林和放射性标记的氟康唑抑制氟康唑的能量依赖性外排，导致真菌细胞中氟康唑的细胞内浓度增加。
Magnaporthe oryzae is an agricultural mold that causes disease in rice, resulting in devastating crop losses. Since rice is a world-wide staple food crop, infection by M. oryzae poses a serious global food security threat. Fungicides, including azole antifungals, are used to prevent and combat M. oryzae plant infections. The target of azoles is CYP51, an enzyme localized on the endoplasmic reticulum (ER) and required for fungal ergosterol biosynthesis. However, many basic drug-pathogen interactions, such as how the azole gets past the fungal cell wall and plasma membrane, and is transported to the ER, are not understood. In addition, reduced intracellular accumulation of antifungals has consistently been observed as a drug resistance mechanism in many fungal species. Studying the basic biology of drug-pathogen interactions may elucidate uncharacterized mechanisms of drug resistance and susceptibility in M. oryzae and potentially other related fungal pathogens. We characterized intracellular accumulation of azole drugs in M. oryzae using a radioactively labeled fluconazole uptake assay to gain insight on whether azoles enter the cell by passive diffusion, active transport, or facilitated diffusion. We show that azole accumulation is not ATP-dependent, nor does it rely on a pH-dependent process. Instead there is evidence for azole drug uptake in M. oryzae by a facilitated diffusion mechanism. The uptake system is specific for azole or azole-like compounds and can be modulated depending on cell phase and growth media. In addition, we found that co-treatment of M. oryzae with ‘repurposed’ clorgyline and radio-labeled fluconazole prevented energy-dependent efflux of fluconazole, resulting in an increased intracellular concentration of fluconazole in the fungal cell.
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