Building a quiet cell cycle clock

构建安静的细胞周期时钟

• 批准号: 8237988
• 负责人: FREDERICK R. CROSS
• 金额: $ 33.9万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-08-01 至 2015-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8237988
• 关键词: Algae; Animals; Behavior; Biological; Biological Models; Biology; Cataloging; Catalogs; Cell Cycle; Cell Cycle Regulation; Cell Size; Cell division; Cells; Chlamydomonas reinhardtii; Commit; Complex; Conflict (Psychology); Coupling; Cyclin A; Cyclins; Data; Detection; Dimensions; Equilibrium; Eukaryotic Cell; Event; Explosion; Family; Fluorescence Microscopy; Funding; Gene Expression; Genes; Genetic Screening; Green Algae; Growth and Development function; Human Development; Image Analysis; Indium; Individual; Learning; Link; M cell; Measures; Methods; Microbial Genetics; Microbiology; Mitosis; Mitotic; Mitotic Cell Cycle; Modeling; Mutation; Nature; Newborn Infant; Noise; Organism; Peripheral; Phase; Phosphoric Monoester Hydrolases; Phosphotransferases; Plants; Process; Proteins; Quantitative Genetics; Reagent; Regulation; Retinoblastoma Protein; Robotics; S Phase; Saccharomycetales; Source; System; Systems Theory; Temperature; Testing; Time; Work; Yeast Model System; Yeasts; base; biological systems; controlled release; cost; human disease; innovation; insight; mathematical model; mutant; protein function; research study; response; success

Cell cycle control is highly conserved through the eukaryotic kingdom, and the budding yeast model system has been the source of major insights applicable to issues in human development and disease. This proposal continues systems-level analysis of the eukaryotic cell cycle using this model system. The cell cycle 'clock' functions with high reliability and low noise, even though individual components and circuits making up the clock are frequently known to be highly variable. For example, gene expression is known to be highly variable between individual cells, and yet cell-cycle-regulated gene expression can be highly reliable with respect to timing and amplitude. Threshold responses to rising cyclin-Cdk activity levels can provide switch-like behavior, but such switches can frequently come at the cost of highly variable onset time; the overall cell cycle control circuitry avoids this variability. We are pursuing an emerging concept of multiple independent oscillators contributing to cell cycle control; while uncoupled oscillators result in highly variable and irregular sequences of cell cycle events, we propose that coupling ('phase-locking') of otherwise independent oscillators to the central cyclin-Cdk oscillator can yield a robust and accurate overall system. This proposal continues our innovative use of quantitative time-lapse fluorescence microscopy, over multi-cell cycle timescales, combined with semi-automated image analysis and in-depth genetic and quantitative analysis to drive systems-level understanding of cell cycle control. We are developing new methods of mathematical modeling. There is a pressing need in biology for simple but experimentally constrained models that can reveal basic control principles. The challenge is to find the most illuminating balance between the detail required for a connection to biological reality, and model simplicity required for transparency and insight. We are exploring methods to use geometrical, low-dimensionality representations of the cell cycle control network that can still be experimentally constrained, and that will yield testable predictions. In a new direction to provide evolutionary contrast from a critical but underexplored branch of the eukaryotic kingdom, we will carry out a genetic screen aiming at saturated detection of cell cycle control elements in the green alga, Chlamydomonas reinhardtii. We have devised robotic methods for the microbiology of mutant isolation, which combined with deep sequencing, allows a massive speedup of this project compared to traditional means.

细胞周期控制通过真核王国高度保守，发芽的酵母 模型系统一直是适用于人类发展问题的主要见解的来源 和疾病。该建议继续使用系统级分析真核细胞周期 这个模型系统。细胞周期“时钟”功能具有很高的可靠性和低噪声，即使 尽管构成时钟的单个组件和电路通常是 高度可变。例如，已知基因表达在个体之间是高度变化 细胞，但相对于时间，细胞周期调节的基因表达可以高度可靠 和振幅。阈值对上升的细胞周期蛋白-CDK活性水平的反应可以提供类似开关的 行为，但是这种开关通常可以以高度可变的发作时间为代价。 总体细胞周期控制电路避免了这种可变性。我们正在追求新兴 有助于细胞周期控制的多个独立振荡器的概念；虽然没有耦合 振荡器导致高度可变和不规则的细胞周期事件序列，我们建议 其他独立振荡器的耦合（“相锁”）与中央细胞周期蛋白-CDK 振荡器可以产生强大而准确的总体系统。该提议继续我们 多细胞周期的定量延时荧光显微镜的创新使用 时间尺度，结合半自动图像分析和深入的遗传和 定量分析以驱动系统级别对细胞周期控制的理解。 我们正在开发新的数学建模方法。生物学需求紧迫 对于可以揭示基本控制原理的简单但实验约束的模型。这 挑战是在连接到与 生物学现实以及透明度和洞察力所需的模型简单性。我们正在探索 使用几何，低维表示网络周期控制网络的方法 这仍然可以通过实验限制，这将产生可测试的预测。在新的 从关键但未充分忽略的分支提供进化对比的方向 真核生物王国，我们将执行旨在饱和检测细胞周期的遗传屏幕 绿色藻类的控制元素，衣原体Reinhardtii。我们设计了机器人 突变隔离的微生物学方法，结合了深度测序，可以 与传统手段相比，该项目的巨大加速。