Discovering Immediate-Early Events in Hedgehog Signal Transduction

描述（由申请人提供）：大多数器官和组织的发育需要信号转导途径，如Hedgehog（Hh）、TGF 1、Notch和Wnt。大多数研究都集中在这些途径的既定组成部分，它们之间的信息流，以及它们驱动的相对缓慢的转录变化。相反，我们提出了一个新的系统的方法来发现重要的快速反应的发展信号，使用Hh信号作为测试案例。具体目标1：我们将确定一个重要的发育信号，刺猬，应用稳定同位素标记与氨基酸在细胞培养（SILAC）和质谱发展调节的即时早期反应。具体目标二：我们将描绘“重新发现”的Hh立即早期反应的组件的功能，并探索新的线索，将提供深入了解信号转导。总结：对主要发育信号的最早反应的全面评估将揭示信号转导机制的新的和关键的方面。我们使用一种途径的测试将为将该方法应用于许多其他途径奠定基础。

DESCRIPTION (provided by applicant): The development of most organs and tissues requires signal transduction pathways such as Hedgehog (Hh), TGF1, Notch, and Wnt. Most studies have focused on established components of these pathways, information flow between them, and the relatively slow transcriptional changes they drive. We propose instead a new systematic approach to discovering important rapid responses to developmental signals, using Hh signaling as a test case. Specific Aim 1: We will identify immediate-early responses to an important developmental signal, Hedgehog, applying Stable Isotope Labeling with Amino acids in Cell culture (SILAC) and mass spectrometry to developmental regulation. Specific Aim 2: We will delineate the functions of "re-discovered" components in the Hh immediate-early response and explore novel leads that will provide insight into signal transduction. Summary: Comprehensive assessment of the earliest responses to major developmental signals will reveal new and critical aspects of the signal transduction mechanisms. Our tests using one pathway will set the stage for applying the approach to many others.