MAPK Signaling in Single Yeast Cells: Dynamics, Variability, and Evolution

单酵母细胞中的 MAPK 信号传导：动力学、变异性和进化

• 批准号: 7917742
• 负责人: ALEXANDER VAN OUDENAARDEN
• 金额: $ 70.8万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2011-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7917742
• 关键词: Apoptosis; Asthma; Autoimmunity; Behavior; Biochemical; Biochemical Genetics; Biological Assay; Biological Models; Biological Process; Candida albicans; Candida glabrata; Cell Nucleus; Cell physiology; Cells; Chromatin; Computer Architectures; Data; Disease; Distant; Elements; Eukaryota; Event; Evolution; Feedback; Gene Expression; Gene Proteins; Genetic; Genetic Transcription; Genomics; Glycerol; Goals; Hybrids; Indium; Individual; Inflammatory; Kinetics; Knock-out; Laboratories; Life; Link; Maps; Measurement; Measures; Mediating; Metabolism; Methods; Mitogen Activated Protein Kinase 1; Mitogen-Activated Protein Kinases; Modeling; Monitor; Nuclear; Nuclear Export; Orthologous Gene; Osmolar Concentration; Osmotic Pressure; Pathway interactions; Pheromone; Phosphoric Monoester Hydrolases; Phosphotransferases; Physiologic pulse; Play; Point Mutation; Population; Process; Property; Proteins; Psychological Techniques; Public Health; Reaction; Regulation; Research Personnel; Resolution; Role; Saccharomyces; Saccharomyces cerevisiae; Saccharomycetales; Series; Signal Pathway; Signal Transduction; Speed; Stimulus; Stress; System; Time; Titrations; Transcription Coactivator; Transplantation; Work; Yeasts; comparative; cytokine; experimental analysis; extracellular; fungus; genome sequencing; insight; predictive modeling; programs; research study; response; tool; transcription factor; upstream kinase

DESCRIPTION (provided by applicant): Mitogen-activated protein kinase (MARK) pathways are evolutionary-conserved eukaryotic signaling modules that regulate diverse cellular processes. In response to a multitude of extracellular stimuli, MAPKs phosphorylate and activate downstream transcription factors that modify chromatin and orchestrate gene expression. Many MARK pathways have been genetically defined in Saccharomyces cerevisiae. In this proposal we focus on one of the well-characterized yeast MARK pathways, the high osmolarity glycerol (HOG) response pathway. During the last few decades, the components and the regulatory network of this cascade have been elucidated using genetic and biochemical assays performed on large populations of cells. In this proposal we take a complementary approach by monitoring the signaling dynamics in single cells with high temporal resolution. This method provides three distinct advantages compared to traditional biochemical and genetic assays on cell populations: (1) MARK activity will be measured in live cells; (2) measurements are almost instantaneous giving a temporal resolution on the order of 1 second which is the typical time-scale of the kinetics of individual reaction steps; (3) measurements will be performed on many single cells in parallel, allowing studies of cell-to-cell variability in signaling dynamics and allowing a direct measurement of the correlation between the concentration of a key component in the network and the signaling dynamics in a single cell. The specific aims are organized around measuring the signaling dynamics at three different time-scales: (1) the initial rapid signal propagation before feedback regulation is activated; (2) adaptation dynamics dominated by feedback regulation and (3) evolutionary time-scales, which will be explored by using a comparative experimental analysis of the HOG signaling pathway of related yeast species. The proposed experimental work will be closely integrated with quantitative modeling approaches with the ultimate goal to build a predictive model of the HOG response pathway. Relevance to public health: Some mammalian MAPKs are activated by inflammatory cytokines and environmental stresses which might play an important role in diseases like asthma and autoimmunity. Because MARK cascades are highly conserved, the proposed experimental and theoretical techniques and concepts that will be important for the analysis of MARK cascades in higher eukaryotes.

描述（由申请人提供）：有丝分裂原激活的蛋白激酶（MARK）途径是调节多种细胞过程的进化保存的真核信号传导模块。为了响应多种细胞外刺激，MAPK磷酸化并激活下游转录因子，以修饰染色质并编排基因表达。在酿酒酵母中，已经在遗传上定义了许多标记途径。在此提案中，我们着重于特征良好的酵母标记途径之一，即高渗透压甘油（HOG）响应途径。在过去的几十年中，使用对大量细胞种群进行的遗传和生化测定法阐明了该级联反应的组件和调节网络。在此提案中，我们通过监视具有高时间分辨率的单个单元的信号传导动力学来采用互补方法。与传统的生化和遗传测定相比，该方法具有三个不同的优势：（1）在活细胞中测量标记活性； （2）测量几乎是瞬时的，给出了1秒钟的时间分辨率，这是单个反应步骤的动力学的典型时间尺度； （3）将在许多单个单元上进行测量，并在许多单个单元格上并行进行信号动力学中的细胞间变异性研究，并可以直接测量网络中关键成分浓度与单个单元格中的信号动力学之间的相关性。具体目的是围绕在三个不同时间尺度下测量信号动力学的围绕：（1）激活反馈调控之前的初始快速信号传播； （2）由反馈调节和（3）进化时间尺度主导的适应动力学，这将通过使用对相关酵母菌物种的猪信号传导途径的比较实验分析来探讨。提出的实验工作将与定量建模方法紧密整合，最终目标是建立猪反应途径的预测模型。与公共卫生有关：一些哺乳动物的MAPK被炎症性细胞因子和环境压力激活，这可能在哮喘和自身免疫性等疾病中起重要作用。由于Mark Cascades是高度保守的，因此提出的实验和理论技术和概念对于分析高等真核生物中的Mark Cascades至关重要。