| Abstract | 第6-7页 |
| 中文摘要 | 第8-10页 |
| Abbreviations | 第10-13页 |
| Chapter Ⅰ:Review | 第13-61页 |
| 1. Hedgehog signal network in Drosophila | 第14-38页 |
| 1.1 Core components of Hedgehog pathway in Drosophila | 第16-24页 |
| 1.1.1 Hedgehog | 第16-17页 |
| 1.1.2 Patched | 第17-18页 |
| 1.1.3 Smoothened | 第18-20页 |
| 1.1.4 Cubitus interruptus | 第20-22页 |
| 1.1.5 Fused | 第22-23页 |
| 1.1.6 Suppressor of Fused | 第23页 |
| 1.1.7 Costal2 | 第23-24页 |
| 1.2 The Hedgehog pathway in vertebrate | 第24-29页 |
| 1.3 Regulation of Hedgehog pathway | 第29-33页 |
| 1.3.1 Phosphorylation | 第30-32页 |
| 1.3.2 Ubiquitination | 第32-33页 |
| 1.3.3 Additional regulations | 第33页 |
| 1.4 Hedgehog-related diseases | 第33-38页 |
| 2. Ubiquitin-proteasome system | 第38-45页 |
| 2.1 The key components of ubiquitin-proteasome system | 第39-43页 |
| 2.1.1 The ubiquitin-activating enzyme,E1 | 第39页 |
| 2.1.2 The ubiquitin-conjugating enzyme,E2 | 第39-40页 |
| 2.1.3 The ubiquitin-protein ligase,E3 | 第40-41页 |
| 2.1.4 The proteasome | 第41-43页 |
| 2.2 Multifaceted modes and roles of ubiquitination | 第43-45页 |
| References | 第45-61页 |
| Chapter Ⅱ:Deubiquitination of Ci/Gli by Usp7/HAUSP regulates Hedgehogsignaling | 第61-117页 |
| 1. Abstract | 第62页 |
| 2. Introduction | 第62-64页 |
| 3. Materials and Methods | 第64-71页 |
| 3.1 Constructs, Mutants, and Transgenes | 第64-65页 |
| 3.2 Immunostaining and in situ hybridization of wing discs | 第65-66页 |
| 3.3 Cell culture, Transfection, Immunoprecipitation, Western blot, Cell immunostaining, Luciferase reporter assay | 第66-67页 |
| 3.4 Generating usp7 mutation clones | 第67页 |
| 3.5 GST fusion protein pull-down assay | 第67-68页 |
| 3.6 RNA interference | 第68页 |
| 3.7 Ci protein stability assays and ubiquitination assay | 第68-69页 |
| 3.8 In situ hybridization of zebrafish embryos | 第69页 |
| 3.9 MO knockdown | 第69页 |
| 3.10 RNA isolation, Reverse transcription, and Real-time PCR | 第69-70页 |
| 3.11 MTT cell proliferation assay | 第70-71页 |
| 3.12 BrdU incorporation assay | 第71页 |
| 4. Results | 第71-108页 |
| 4.1 Loss of usp7 specifically compromises Hh signaling through promoting Ci protein degradation | 第71-78页 |
| 4.1.1 Loss of usp7 downregulates Hh signaling in Drosophila | 第71-75页 |
| 4.1.2 Loss of usp7 downregulates Ci through promoting Ci degradation | 第75-77页 |
| 4.1.3 Loss of usp7 does not affect other signaling pathways | 第77-78页 |
| 4.2 Usp7 interacts with Ci | 第78-84页 |
| 4.2.1 Usp7 binds Ci through N-terminal MATH domain | 第78-80页 |
| 4.2.2 Ci binds Usp7 through its multiple P/AxxS motifs | 第80-83页 |
| 4.2.3 The interaction of Ci and Usp7 is not regulated by Ci ubiquitination | 第83-84页 |
| 4.3 Hh promotes Ci interaction with Usp7 | 第84-87页 |
| 4.4 Usp7 antagonizes Ci degradation caused by both Slimb-Cull and Hib-Cul3 E3 ligases | 第87-91页 |
| 4.5 Usp7 deubiquitinase activity is essential for Ci stabilization | 第91-93页 |
| 4.6 Usp7 counteracts both Slimb-Cull and Hib-Cul3-dependent ubiquitination of Ci | 第93-94页 |
| 4.7 Usp7 regulates Ci through Usp7-GMPS complex | 第94-97页 |
| 4.8 HAUSP deubiquitinates Gli and promotes Hh signaling activity in mammalian systems | 第97-105页 |
| 4.8.1 HAUSP could functionally replace Usp7 in the regulation of Ci | 第97-98页 |
| 4.8.2 HAUSP binds Gli proteins in a manner promoted by Hh treatment | 第98-100页 |
| 4.8.3 HAUSP stabilizes Gli proteins | 第100-101页 |
| 4.8.4 HAUSP inhibits the ubiquitination of Gli proteins | 第101-103页 |
| 4.8.5 HAUSP positively regulates Hh pathway activity in mammalian cells | 第103-105页 |
| 4.9 zUsp7 plays a conserved positive role in the regulation of Hh signaling in zebrafish | 第105-106页 |
| 4.10 HAUSP promotes the proliferation of Hh-related cancer cells | 第106-108页 |
| 5. Discussion | 第108-111页 |
| References | 第111-117页 |
| Acknowledgements | 第117-118页 |
| Publications | 第118-120页 |
