Mechanisms of Compartmentalized cAMP Signaling

项目总结： 环磷酸腺苷(CAMP)及其效应分子如cAMP依赖蛋白的信号转导 激酶(PKA)和cAMP激活的交换蛋白(EPAC)，调节多种细胞 功能包括细胞生长、增殖、新陈代谢、存活和运动，以及胰岛素 胰腺细胞的分泌物。我们研究的总体目标是阐明 时空调控实现高特异性的分子机制和功能作用 在营地发信号。CAMP信号通路的异常对临床情况有影响 例如肥胖和2型糖尿病，特别是在细胞功能的背景下。一个 从机制上理解cAMP信号的特异性是制定治疗策略的关键 在这些临床情况下。 CAMP效应的空间划分的概念是20年前提出的，但只是 近年来，在细胞环境中研究cAMP信号的创新方法已经成为 可以提供直接的机械证据。然而，尽管最近取得了这些进展，但仍有 在我们对时空调控机制的理解上仍然存在很大差距 坎普和它的效应器。此外，人们对信令信息是如何编码的知之甚少 活动的时空模式被转化为特定的功能反应。 在我们的初步研究中，我们开发了新的分子工具来监测和扰动 活细胞内cAMP/PKA活性的时空分辨率和精确度进一步提高。 此外，基于我们最近在MIN6细胞中发现的钙-cAMP-PKA振荡电路， 我们发现，A-Kinase锚定蛋白79/150(AKAP79/150)组装了一个含有PKA的 这些细胞中的信号复合体，并影响钙-cAMP-PKA通路的活性动态。在……里面 目前的建议，利用我们的新分子工具，并结合计算模型和 实验方法，我们将检验我们的假设，即钙-cAMP-PKA振荡电路， 受AKAPs在空间和时间上的进一步调节，使PKA获得高度的信号特异性 以及通过频率调制实现分集。具体目标是：1)开发新的分子 研究活细胞内cAMP/PKA信号时空调控的工具；2)阐明 Ca~(2+)-cAMP-PKA振荡回路的空间区划和频率控制。

Project summary: Signaling through cyclic AMP (cAMP) and its effector molecules, such as cAMP-dependent protein kinase (PKA) and exchange proteins activated by cAMP (Epac), regulates a variety of cellular functions including cell growth, proliferation, metabolism, survival and mobility, as well as insulin secretion in the case of pancreatic  cells. The overall goal of our research is to elucidate the molecular mechanisms and functional roles of spatiotemporal regulation in achieving high specificity in cAMP signaling. Aberrations in the cAMP signaling pathway have implications for clinical conditions such as obesity and type 2 diabetes mellitus, particularly in the context of -cell functions. A mechanistic understanding of cAMP signaling specificity is crucial to developing therapeutic strategies for these clinical conditions. The concept of spatial compartmentalization of cAMP effects was proposed 20 years ago, but only in recent years have innovative approaches to studying cAMP signaling in the cellular context become available to provide direct mechanistic evidence. However, despite these recent advances, there are still large gaps in our understanding about the mechanisms underlying the spatiotemporal regulation of cAMP and its effectors. Furthermore, little is known about how the signaling information encoded in the spatiotemporal patterns of activities is translated into specific functional responses. In our preliminary studies, we have developed new molecular tools to monitor and perturb cAMP/PKA activities in living cells with further enhanced spatiotemporal resolution and precision. Furthermore, building on our recent discovery of a Ca2+-cAMP-PKA oscillatory circuit in MIN6  cells, we showed that A-Kinase Anchoring Protein 79/150 (AKAP79/150) assembles a PKA-containing signaling complex in these cells and influences the activity dynamics of the Ca2+-cAMP-PKA circuit. In the current proposal, utilizing our new molecular tools and by combining computational modeling and experimental approaches, we will test our hypothesis that the Ca2+-cAMP-PKA oscillatory circuit, further regulated spatially and temporally by AKAPs, allows PKA to achieve high signaling specificity and diversity through frequency modulation. The specific aims are: 1) developing novel molecular tools to interrogate the spatiotemporal regulation of cAMP/PKA signaling in living cells; 2) elucidating the spatial compartmentalization and frequency control of the Ca2+-cAMP-PKA oscillatory circuit.