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.

项目摘要 当从环境中吸入孢子时，许多侵入性真菌疾病开始 并开始主机内的积极增长。尽管它们对真菌生命周期至关重要 作为抗真菌靶标的潜力，孢子休眠的建立和维护是 理解不佳。 拟议的人类病原体曲霉曲霉的试点研究重点是真菌 在休眠的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）和积极生长的细胞中的低水平。具有这种特殊表达模式的基因 指定的HIC（分生孢子高）基因可能参与机构或 维持孢子休眠。在拟议的试点研究中，25个HIC基因 处于休眠与非不介出细胞中的差异转录水平将通过 删除和标记。删除菌株将分析开发，休眠状况的变化 使用流式细胞术和显微镜发芽。标记应变将进行分析 使用荧光发育，休眠和发芽期间HIC蛋白的定位 显微镜。标记的菌株也将用于亲和力纯化和质谱 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对 在真菌分生孢子中建立和维持休眠以及本地化和相互作用 在发育期间和从休眠到积极生长的过程中过渡到休眠的数据。