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 许多侵入性真菌疾病开始时，孢子吸入从环境打破休眠 并在宿主体内开始活跃生长。尽管它们对真菌的生命周期至关重要 和潜在的抗真菌靶标，孢子休眠的建立和维持是 不太了解。 拟议的人类病原体烟曲霉的初步研究重点是真菌- 在休眠的无性孢子中具有高转录水平的功能未知的特定基因 （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（高分生孢子）基因，可能参与建立或 孢子休眠的维持。在拟议的试点研究中，25个hic基因与最 休眠细胞与非休眠细胞中的差异转录物水平将通过以下来表征： 删除和标记。将分析缺失菌株的发育、休眠、 和发芽的能力。将分析标记菌株的 利用荧光定位Hic蛋白在发育、休眠和萌发过程中的位置 显微镜标记的菌株也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， 真菌分生孢子沿着休眠的建立和维持，以及定位和相互作用 发育期间向休眠过渡以及从休眠向活跃生长过渡的数据。