Signaling iteractions in cytokinesis, cell wall biogenesis, and growth in fungi

真菌胞质分裂、细胞壁生物发生和生长中的信号交互

• 批准号: 8132355
• 负责人: JOSE R RODRIGUEZ-MEDINA
• 金额: $ 25.73万
• 依托单位: UNIVERSITY OF PUERTO RICO MED SCIENCES
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-08-01 至 2013-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8132355
• 关键词: Actomyosin; Affect; Antifungal Agents; Biochemical; Biogenesis; Biological Process; Cell Survival; Cell Wall; Cell membrane; Cells; Cellular Stress; Characteristics; Complement; Complex; Cytokinesis; Data; Defect; Development; Down-Regulation; Exhibits; Gene Expression; Genes; Genetic; Genetic Transcription; Goals; Growth; Interruption; Knowledge; Lead; Link; Messenger RNA; Metabolism; Mutation; Myosin ATPase; Myosin Type II; Overlapping Genes; Pathway interactions; Phenotype; Process; Proteins; Ribosomal Proteins; Saccharomyces cerevisiae; Signal Pathway; Signal Transduction; Signal Transduction Pathway; Stress; Testing; Transcriptional Activation; Transducers; Translational Regulation; Yeasts; biological adaptation to stress; cell growth; fungus; gene repression; mRNA Expression; mutant; research study; response; sensor

DESCRIPTION (provided by applicant): The scientific goals of this application are to 1- understand signaling interactions connecting myosin II functions in cytokinesis, cell wall biogenesis, and cell growth 2- understand the relation between myosin II function and cell wall integrity and 3- identify new potential targets for antifungal drugs. We will accomplish this by identifying what metabolic processes are affected by the interruption of cytokinesis by actomyosindependent and actomyosin-independent mutations, what signaling pathways are necessary for cell survival under these conditions, and the biochemical components linking these pathways in these genetic mutants. Myosin ll-deficient Saccharomyces cerevisiae cells (myolA) exhibit multiple phenotypes with characteristics of both cytokinesis and cell wall mutants. To determine if these phenotypes were related to changes in gene expression, we determined the global transcription profile of myol A strains. These studies corroborated that the PKC1-dependent signaling pathway coordinately activates the transcription of cell wall stress genes in these strains. Coordinated down-regulation of ribosomal protein genes also occurs and it was observed that over expression of ribosomal protein genes RPL30 and RPS31 partially restored cell wall function in a myol A strain. Our main hypothesis is that transcriptional activation of cell wall stress genes and the down-regulation of ribosomal protein genes in myol A strains represent separate phenotypes for cell wall and cytokinesis mutants, respectively; each employing distinct essential signaling pathways that are interconnected. To test this hypothesis our specific aims are to: 1- conduct mRNA transcription analysis of the cytokinesis mutant chs2A and the cell wall mutant fksl A for subsequent comparison with myolA strains, 2-conduct analysis of mRNA translational regulation in cytokinesis mutants, 3- determine the status of the PKC1, TOR1, and TOR2 signaling pathways in cytokinesis mutant strains and test their requirement for cell viability, and 4- identify common regulators of the PKC1 and TOR signaling pathways in cytokinesis and cell wall mutants. The significance of this study lies in the discovery of new regulatory connections that coordinate actomyosin function and cell wall biogenesis in fungal cells. The proposed study will also increase our understanding about how loss of myosin II contributes to the stress response in yeast cells. Knowledge of the interaction between myosin II and stress signaling can lead to identification of potential targets for new antifungal drugs.

描述（由申请人提供）：本申请的科学目标是：1-了解肌球蛋白II在细胞分裂、细胞壁生物发生和细胞生长中的信号相互作用；2-了解肌球蛋白II功能和细胞壁完整性之间的关系；3-确定抗真菌药物的新潜在靶点。我们将通过确定哪些代谢过程受到肌动球蛋白不依赖和肌动球蛋白不依赖突变的细胞分裂中断的影响，哪些信号通路是这些条件下细胞存活所必需的，以及这些基因突变中连接这些通路的生化成分来实现这一目标。肌球蛋白缺陷型酿酒酵母细胞（myolA）表现出多种表型，具有细胞质分裂和细胞壁突变的特征。为了确定这些表型是否与基因表达的变化有关，我们确定了myol A菌株的全球转录谱。这些研究证实了pkc1依赖的信号通路在这些菌株中协同激活细胞壁应激基因的转录。核糖体蛋白基因也发生协同下调，观察到核糖体蛋白基因RPL30和RPS31的过表达部分恢复了myol a菌株的细胞壁功能。我们的主要假设是，myol A菌株细胞壁应激基因的转录激活和核糖体蛋白基因的下调分别代表细胞壁突变体和细胞质突变体的不同表型；每一种都采用不同的相互联系的基本信号通路。为了验证这一假设，我们的具体目标是：1-对细胞质分裂突变体chs2A和细胞壁突变体fksl A进行mRNA转录分析，以便与myolA菌株进行后续比较；2-对细胞质分裂突变体进行mRNA翻译调控分析；3-确定细胞质分裂突变株中PKC1、TOR1和TOR2信号通路的状态，并测试其对细胞活力的需求；4-鉴定细胞质分裂和细胞壁突变体中PKC1和TOR信号通路的共同调控因子。本研究的意义在于发现了真菌细胞中协调肌动球蛋白功能和细胞壁生物发生的新的调控联系。拟议的研究也将增加我们对肌球蛋白II的损失如何促进酵母细胞应激反应的理解。了解肌球蛋白II和应激信号之间的相互作用可以确定新的抗真菌药物的潜在靶点。