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Systematic functional analysis of the zinc cluster transcription factor family of the pathogenic yeast Candida albicans by artificial activation

人工激活致病酵母白色念珠菌锌簇转录因子家族的系统功能分析

基本信息

批准号：
201319072
负责人：
Professor Dr. Joachim Morschhäuser
金额：
--
依托单位：
Institut für Molekulare Infektionsbiologie (IMIB)
依托单位国家：
德国
项目类别：
Research Grants
财政年份：
2011
资助国家：
德国
起止时间：
2010-12-31 至 2019-12-31
项目状态：
已结题

来源：
https://gepris.dfg.de/gepris/projekt/201319072?language=en
关键词：
Systematic functional analysis zinc cluster

项目摘要

The regulation of gene expression is crucial for the ability of cells to adapt to changes in their environment, but it is also an important determinant in the development of organisms and in the evolution of species. The zinc cluster proteins are a family of transcriptional regulators that are unique to the fungal kingdom. In the pathogenic yeast Candida albicans they have key roles in the regulation of virulence-related traits. Furthermore, gain-of-function mutations in these transcription factors result in the generation of variants with novel phenotypes and are responsible for the acquisition of antifungal drug resistance in many clinical isolates. However, the functions of most members of this family in the biology and pathogenicity of C. albicans are still unknown. We have devised a method for artificially activating zinc cluster proteins, an approach that does not require knowledge about inducing conditions and can reveal their involvement in a specific phenotype even when deletion of the corresponding genes has no effect. We generated a comprehensive library of C. albicans strains expressing constitutively active forms of all 82 zinc cluster transcription factors of this fungus. The phenotypic analysis of this strain collection identified a novel regulator of the morphological transition to invasive hyphal growth and uncovered previously unknown mechanisms of resistance to the widely used antifungal drug fluconazole and against innate host defenses. Importantly, natural gain-of-function mutations in these transcription factors were detected in patient isolates with the same phenotypes, demonstrating the clinical relevance of our findings. By generating hybrid transcription factors with altered DNA-binding properties, we showed that the ability to efficiently induce the expression of multidrug efflux pumps in the presence of fluconazole strongly increases the intrinsic resistance of C. albicans to this drug. The aims of this project are now to uncover the molecular mechanisms by which specific transcription factors affect the properties and behavior of C. albicans and how important they are for the colonization and infection of different host niches. In addition, we investigate if acquiring the ability to react to a certain stimulus by inducing the expression of specific sets of previously unresponsive genes confers a selective advantage under adverse conditions and contributes to a better adaptation of C. albicans to environmental alterations in the human host.

基因表达的调节对于细胞适应环境变化的能力至关重要，但它也是生物体发育和物种进化的重要决定因素。锌簇蛋白是真菌界特有的转录调节因子家族。在致病性酵母菌白色念珠菌中，它们在调节毒力相关性状中发挥关键作用。此外，这些转录因子的功能获得性突变导致产生具有新表型的变体，并导致许多临床分离株获得抗真菌药物耐药性。然而，该家族的大多数成员在C.白色念珠菌仍然未知。我们设计了一种人工激活锌簇蛋白的方法，这种方法不需要了解诱导条件，即使删除相应的基因没有影响，也可以揭示它们参与特定表型。我们生成了一个全面的C库。表达该真菌的所有82个锌簇转录因子的组成型活性形式的白色念珠菌菌株。该菌株的表型分析确定了一种新的调节剂的形态过渡到侵入性菌丝生长，并发现了以前未知的耐药机制，广泛使用的抗真菌药物氟康唑和先天宿主防御。重要的是，在具有相同表型的患者分离株中检测到这些转录因子的天然功能获得性突变，证明了我们的发现的临床相关性。通过产生具有改变的DNA结合特性的杂交转录因子，我们表明，在氟康唑存在下有效诱导多药外排泵表达的能力强烈增加了念珠菌的内在耐药性。白色念珠菌对这种药物。本项目的目的是揭示特定转录因子影响C.白念珠菌和他们是如何重要的殖民和感染不同的主机壁龛。此外，我们调查，如果获得的能力，以应对某种刺激诱导表达的特定组以前无反应的基因赋予在不利条件下的选择优势，并有助于更好地适应C。白色念珠菌对人类宿主环境的改变。