Investigation of uncharacterized fungal-specific genes in dormant conidia

休眠分生孢子中未表征的真菌特异性基因的研究

• 批准号: 10648082
• 负责人: MICHELLE MOMANY
• 金额: $ 7.55万
• 依托单位: UNIVERSITY OF GEORGIA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-01-18 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10648082
• 关键词: Affinity Chromatography; Antifungal Agents; Aspergillus fumigatus; Cells; Collection; Data; Development; Disease; Environment; Flow Cytometry; Fluorescence Microscopy; Fungal Spores; Future; Gene Deletion; Genes; Germination; Growth; Human; Inhalation; Investigation; Knock-out; Life Cycle Stages; Lung; Maintenance; Mass Spectrum Analysis; Metabolism; Microscopy; Pattern; Persons; Pilot Projects; Predisposition; Proteins; Reproduction spores; Resistance; Stress; Time; Transcript; Yeasts; asexual; differential expression; fungus; gene product; human pathogen; insight; pathogenic fungus

Project Summary Many invasive fungal diseases start when spores inhaled from the environment break dormancy and begin active growth within the host. Despite their critical importance to the fungal life cycle and potential as antifungal targets, the establishment and maintenance of spore dormancy are poorly understood. The proposed pilot study in the human pathogen Aspergillus fumigatus focuses on fungal- specific genes of unknown function that have high transcript levels in dormant asexual spores (conidia) and low levels in actively growing cells. Genes with this particular expression pattern, designated hic (high in conidia) genes, are likely to be involved in the establishment or maintenance of spore dormancy. In the proposed pilot study, the 25 hic genes with the most differential transcript levels in dormant vs nondormant cells will be characterized through deletion and tagging. Deletion strains will be analyzed for changes in development, dormancy, and germination using flow cytometry and microscopy. Tagged strains will be analyzed for localization of Hic proteins during development, dormancy, and germination using fluorescence microscopy. Tagged strains will also be used in affinity purification and mass spectroscopy to identify interacting proteins. Successful completion of the proposed pilot study will identify a set of genes important for establishing and maintaining dormancy in fungal conidia along with localization and interaction data for the transition to dormancy during development and from dormancy to active growth.

项目概要 当从环境中吸入的孢子打破休眠时，许多侵袭性真菌病就会开始 并开始在宿主体内积极生长。尽管它们对真菌生命周期至关重要 以及作为抗真菌靶点的潜力，孢子休眠的建立和维持是 不太了解。 拟议的针对人类病原体烟曲霉的初步研究重点是真菌- 休眠无性孢子中具有高转录水平的未知功能的特定基因 （分生孢子）和活跃生长细胞中的低水平。具有这种特殊表达模式的基因， 指定的 hic（分生孢子含量高）基因，可能参与建立或 维持孢子休眠。在拟议的试点研究中，最常见的 25 个 hic 基因 休眠细胞与非休眠细胞的转录水平差异将通过以下方式表征 删除和标记。将分析缺失菌株的发育、休眠、 并使用流式细胞术和显微镜进行发芽。将分析标记的菌株 使用荧光定位发育、休眠和萌发过程中的 Hic 蛋白 显微镜。标记的菌株还将用于亲和纯化和质谱分析 识别相互作用的蛋白质。 成功完成拟议的试点研究将确定一组重要的基因 建立和维持真菌分生孢子的休眠以及定位和相互作用 发育过程中向休眠过渡以及从休眠到活跃生长的数据。