Fungal Zn(II)(2)Cys(6) Transcription Factor ADS-1 Regulates Drug Efflux and Ergosterol Metabolism under Antifungal Azole Stress.
Fungal Zn(II)(2)Cys(6) Transcription Factor ADS-1 Regulates Drug Efflux and Ergosterol Metabolism under Antifungal Azole Stress.
复制标题
真菌 Zn(II)(2)Cys(6) 转录因子 ADS-1 在抗真菌唑应激下调节药物流出和麦角甾醇代谢。
DOI：
10.1128/aac.01316-20
复制
发表时间：
2021
影响因子：
4.9
通讯作者：
Li Shaojie
中科院分区：
文献类型：
--
作者：
Yin Yajing;Zhang Hanxing;Zhang Yu;Hu Chengcheng;Sun Xianyun;Liu Wei;Li Shaojie
唑类抗真菌药物是临床和农业实践中应用最广泛的抗真菌药物。真菌可以通过改变许多基因的转录水平来适应唑类胁迫，而唑类胁迫的应答机制是真菌产生唑类抗性的基础。在这项研究中，我们确定了一个新的锌（II）2Cys 6转录因子，ADS-1，具有积极的调节功能，在转录反应唑胁迫模式丝状真菌物种粗糙脉孢菌。在酮康唑（KTC）胁迫下，N.粗鲁。ads-1的缺失增加了对不同唑类药物的敏感性，而其过表达增加了对这些唑类药物的耐药性。编码唑类药物关键外排泵的ecdr 4基因受ADS-1的正调控。ads-1的缺失降低了cdr 4对KTC胁迫的转录响应，增加了KTC胁迫下细胞内KTC的积累，而ads-1的过表达则相反。ADS-1还正调控了对KTC胁迫的转录响应byerg 11，byerg 11编码麦角甾醇生物合成的唑靶羊毛甾醇14α-脱甲基酶。经KTC处理后，theads-1缺失突变体的麦角甾醇含量比野生型低，但羊毛甾醇含量比野生型高，而theads-1过表达突变体的麦角甾醇含量比野生型高。ADS-1的同源物广泛存在于子囊菌门的丝状真菌物种中，但不存在于酵母中。黄曲霉中编码ADS-1同源物的基因缺失也增加了对KTC和伊曲康唑（ITZ）的敏感性。此外，A. flavusads-1（Afads-1）显著降低了A.黄曲霉对KTC胁迫的耐受性较强，缺失突变体积累了更多的KTC，但麦角甾醇含量较低。综上所述，这些研究结果表明，ADS-1同源物在不同真菌物种中的唑类反应和唑类的基础抗性中的功能和调节机制是高度保守的。
Antifungal azoles are the most widely used antifungal drugs in clinical and agricultural practice. Fungi can mount adaptive responses to azole stress by modifying the transcript levels of many genes, and the responsive mechanisms to azoles are the basis for fungi to develop azole resistance. In this study, we identified a new Zn(II)2Cys6transcription factor, ADS-1, with a positive regulatory function in transcriptional responses to azole stress in the model filamentous fungal species Neurospora crassa. Under ketoconazole (KTC) stress, theads-1transcript level was significantly increased in N. crassa. Deletion ofads-1increased susceptibility to different azoles, while its overexpression increased resistance to these azoles. Thecdr4gene, which encodes the key azole efflux pump, was positively regulated by ADS-1. Deletion ofads-1reduced the transcriptional response bycdr4to KTC stress and increased cellular KTC accumulation under KTC stress, whileads-1overexpression had the opposite effect. ADS-1 also positively regulated the transcriptional response byerg11, which encodes the azole target lanosterol 14α-demethylase for ergosterol biosynthesis, to KTC stress. After KTC treatment, theads-1deletion mutant had less ergosterol but accumulated more lanosterol than the wild type, whileads-1overexpression had the opposite effect. Homologs of ADS-1 are widely present in filamentous fungal species of Ascomycota but not in yeasts. Deletion of the gene encoding an ADS-1 homolog in Aspergillus flavus also increased susceptibility to KTC and itraconazole (ITZ). Besides, deletion of A. flavusads-1(Afads-1) significantly reduced the transcriptional responses by genes encoding homologs of CDR4 and ERG11 in A. flavus to KTC stress, and the deletion mutant accumulated more KTC but less ergosterol. Taken together, these findings demonstrate that the function and regulatory mechanism of ADS-1 homologs among different fungal species in azole responses and the basal resistance of azoles are highly conserved.