Discovering Immediate-Early Events in Hedgehog Signal Transduction

描述(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)、TGF1、Notch和Wnt。大多数研究都集中在这些通路的既定组成部分，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，我们提出了一种新的系统方法来发现对发育信号的重要快速反应，并将HH信号作为测试案例。具体目标1：我们将应用细胞培养中氨基酸稳定同位素标记(SILAC)和质谱学技术来鉴定对重要发育信号Hedgehog的即刻-早期反应。具体目标2：我们将描述HH即刻-早期反应中“重新发现”的成分的功能，并探索将为信号转导提供洞察力的新线索。摘要：对主要发育信号最早反应的综合评估将揭示信号转导机制的新的和关键的方面。我们使用一条途径进行的测试将为将该方法应用于其他许多途径奠定基础。

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.