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都需要Hedgehog （Hh）、TGF1、Notch、Wnt等信号转导通路。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。我们提出了一种新的系统方法来发现对发育信号的重要快速反应，使用Hh信号作为测试案例。具体目标1：我们将利用细胞培养中氨基酸稳定同位素标记（SILAC）和质谱技术来确定对重要发育信号刺猬的即时早期反应。具体目标2：我们将描述Hh即时早期反应中“重新发现”的成分的功能，并探索新的线索，为信号转导提供见解。摘要：对主要发育信号的早期反应的综合评估将揭示信号转导机制的新和关键方面。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.