Inhibiting fungal multidrug resistance by disrupting an activator-Mediator interaction.
Inhibiting fungal multidrug resistance by disrupting an activator-Mediator interaction.
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DOI：
10.1038/nature16963
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发表时间：
2016-02-25
期刊：
影响因子：
64.8
通讯作者：
Arthanari H
中科院分区：
文献类型：
--
作者：
Nishikawa JL;Boeszoermenyi A;Vale-Silva LA;Torelli R;Posteraro B;Sohn YJ;Ji F;Gelev V;Sanglard D;Sanguinetti M;Sadreyev RI;Mukherjee G;Bhyravabhotla J;Buhrlage SJ;Gray NS;Wagner G;Näär AM;Arthanari H
真核转录激活剂通过招募共激活剂（例如 RNA 聚合酶 II 相互作用介体复合物）来刺激特定靶基因组的表达。转录激活子的异常功能与多种疾病有关。然而，由于缺乏对疾病相关转录激活剂基因激活机制的详细分子知识，靶向治疗工作受到了阻碍。我们之前在人类 MED15 介体亚基中发现了一个激活剂靶向的三螺旋束 KIX 结构域，该结构域在真菌的 Gal11/Med15 介体亚基中结构保守。 Gal11/Med15 KIX 结构域与多效耐药转录因子 (Pdr1) 直向同源物结合，后者是酿酒酵母和临床上重要的人类病原体光滑念珠菌多药耐药途径的关键调节因子。 C.流行程度glabratais 上升，部分原因是其对唑类（最广泛使用的抗真菌剂）的内在敏感性较低。 C. 耐药临床分离株glabrata 最常在 Pdr1 中包含点突变，使其具有组成型活性，，，，，，表明该转录激活途径代表了 C 中的关键。光滑菌多重耐药性。在这里，我们进行连续的生化和体内高通量筛选，以鉴定 C 相互作用的小分子抑制剂。 glabrataPdr1 激活域与 C. glabrataGal11A KIX 结构域。先导化合物 (iKIX1) 抑制 Pdr1 依赖性基因激活并使耐药 C 重新敏感。光滑唑类抗真菌药，在体外和动物模型中用于播散性和泌尿道 C.光滑感染。确定C的NMR结构。 glabrataGal11A KIX 结构域提供了对 Pdr1 基因激活和 iKIX1 抑制多药耐药性的分子机制的详细了解。我们已经证明了介导物中转录因子结合位点的小分子靶向作为真菌感染性疾病的新型治疗策略的可行性。
Eukaryotic transcription activators stimulate the expression of specific sets of target genes through recruitment of co-activators such as the RNA polymerase II-interacting Mediator complex,. Aberrant function of transcription activators has been implicated in several diseases. However, therapeutic targeting efforts have been hampered by a lack of detailed molecular knowledge of the mechanisms of gene activation by disease-associated transcription activators. We previously identified an activator-targeted three-helix bundle KIX domain in the human MED15 Mediator subunit that is structurally conserved in Gal11/Med15 Mediator subunits in fungi,. The Gal11/Med15 KIX domain engages pleiotropic drug resistance transcription factor (Pdr1) orthologues, which are key regulators of the multidrug resistance pathway inSaccharomyces cerevisiaeand in the clinically important human pathogenCandida glabrata,. The prevalence ofC. glabratais rising, partly owing to its low intrinsic susceptibility to azoles, the most widely used antifungal agent,. Drug-resistant clinical isolates ofC. glabratamost commonly contain point mutations in Pdr1 that render it constitutively active,,,,,, suggesting that this transcriptional activation pathway represents a linchpin inC. glabratamultidrug resistance. Here we perform sequential biochemical andin vivohigh-throughput screens to identify small-molecule inhibitors of the interaction of theC. glabrataPdr1 activation domain with theC. glabrataGal11A KIX domain. The lead compound (iKIX1) inhibits Pdr1-dependent gene activation and re-sensitizes drug-resistantC. glabratato azole antifungalsin vitroand in animal models for disseminated and urinary tractC. glabratainfection. Determining the NMR structure of theC. glabrataGal11A KIX domain provides a detailed understanding of the molecular mechanism of Pdr1 gene activation and multidrug resistance inhibition by iKIX1. We have demonstrated the feasibility of small-molecule targeting of a transcription factor-binding site in Mediator as a novel therapeutic strategy in fungal infectious disease.