Unraveling the quantitative dynamics of cAMP-PKA signaling in yeast

项目摘要 从细胞的蓝图开始，包括其信号电缆的地图，我们现在面对的是 了解信号如何在这些电缆中流动并破译的巨大挑战 蜂窝信息传输协议。 一个特别令人生畏的问题是，细胞经常使用共享的路径（相同的电缆）， 传输不同的信号，但有精确的能力来具体解释这些信号。 细胞如何编码信息，然后以高保真度对其进行解码，以优化共享的 信号传导渠道在很大程度上仍是未知的。发现这些战略需要协同作用 在独特的多学科技术工具包、计算建模和高 分辨率/高通量实验研究。使用蛋白激酶A（PKA）， 重要的生物信号枢纽，和我们独特的多学科方法，我们将剖析 不同的输入被S.酿酒酵母和 然后由下游转录因子和基因解码。我们将调查 这样的信号编码和解码方案中的错误的影响，并且通过这样做，我们 将能够提取细胞用来规避其信息的基本原则 传输瓶颈。从根本上说，这些调查将提供一个全面的 并定量描绘PKA通路作为一个完整系统的动态运作， 以及其失效模式及其与细胞生理学的联系的详细概要。我们 实验设计还将产生PKA与其他蛋白质的连接目录。 蜂窝底盘

PROJECT SUMMARY Starting from a blueprint of the cell, including a map of its signaling cables, we now face the monumental challenge of understanding how signals flow in these cables and deciphering cellular information transmission protocols. One particularly daunting problem is that cells often use shared pathways (the same cables) to transmit different signals, yet have an exquisite capacity to specifically interpret these signals. How cells encode information and then decode it with high fidelity to optimize the use of shared signaling channels remains largely unknown. Uncovering these strategies requires synergy between a unique multidisciplinary toolkit of technology, computational modeling, and high resolution/high throughput experimental investigations. Using Protein Kinase A (PKA), an important biological signaling hub, and our unique multidisciplinary approach, we will dissect the mechanisms by which different inputs are encoded by the PKA channel in S. cerevisiae and then decoded by downstream transcription factors and genes. We will investigate the repercussions of errors in such signal encoding and decoding schemes, and by doing so, we will be able to extract fundamental principles that cells use to circumvent their information transmission bottlenecks. Fundamentally, these investigations will provide a comprehensive and quantitative portrait of the dynamic operation of the PKA pathway as an integrated system, and a detailed synopsis of its failure modes and their connections to cellular physiology. Our experimental design will also generate a catalogue of the connectivity of PKA to the rest of the cellular chassis.