Hedgehog Signal Transduction Across the Cell Membrane

跨细胞膜的刺猬信号转导

• 批准号: 7575224
• 负责人: LAWRENCE LUM
• 金额: $ 29.35万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-02-01 至 2011-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7575224
• 关键词: Basal cell carcinoma; Biological Assay; Cell membrane; Cells; Classification; Complex; Cultured Cells; Cytoplasmic Protein; Cytoplasmic Tail; Deformity; Detection; Development; Diagnosis; Disease; Disease Progression; Double-Stranded RNA; Drosophila genome; Drosophila genus; Erinaceidae; Face; Genes; Goals; Heparan Sulfate Proteoglycan; Immunoglobulins; Integral Membrane Protein; Intracellular Membranes; Libraries; Malignant Neoplasms; Malignant neoplasm of pancreas; Malignant neoplasm of prostate; Maps; Measures; Mediating; Membrane; Modeling; Mus; Pathway interactions; Phosphorylation; Post Translational Modification Analysis; Prevention; Process; Prosencephalon; Protein Family; Proteins; RNA library; Reagent; Recruitment Activity; Regulation; Signal Pathway; Signal Transduction; Technology; Testing; Therapeutic Intervention; base; cell growth; extracellular; gain of function; gene function; hedgehog signal transduction; human SMO protein; insight; member; mouse genome; novel; protein protein interaction; receptor; receptor function; response; tool; transcription factor; transmission process

The Hedgehog (Hh) family of proteins coordinates cellular growth and differentiation in a number of developmental contexts by eliciting graded responses in the cells surrounding Hh-producing cells. Aberrant Hh pathway activity is associated with developmental deformities of the face and forebrain, and prevalent cancers such as prostate cancer and basal cell carcinoma (BCC). Understanding how the Hh signal is conveyed from the extracellular milieu to intracellular effectors is therefore pivotal to the successful prevention, detection, and treatment of these and other diseases associated with Hh signaling. This proposal focuses on the mechanisms that allow cellular recognition of the secreted Hh protein at the plasma membrane and subsequent transduction of a signal across the membrane to induce specific intracellular responses. Direct interaction of Hh with the twelve transmembrane protein Patched (Ptc) is required for relinquishing Ptc-mediated suppression of the seven transmembrane protein Smoothened (Smo) in target cells. This interaction is facilitated by Dally-like protein (Dip), a heparan sulfate proteoglycan, and CG9211, a member of the immunoglobulin (Ig) superfamily of receptors. Activated Smo transduces a signal across the membrane in a process that involves recruiting to its cytoplasmic tail a large regulatory complex that includes the transcription factor Cubitus interruptus (Ci). The mechanisms by which Dip, CG9211, and these cytoplasmic protein-protein interactions contribute to Hh pathway response are unclear. In order to study reception and transmission of the Hh signal at the cell membrane, we have developed novel tools that include cultured cell-based assays for pathway responsiveness, reagents for isolating and detecting Hh signaling complexes, and double-stranded RNA libraries that inhibit specific gene function in Drosophila and mouse by RNA-mediated interference (RNAi). By incorporating these tools in a biochemically- and genetically-based strategy, we propose to: 1) determine how Hh signal is sensed at the cell membrane, 2) determine how Smo transduces the Hh signal to cytoplasmic components, and 3) identify Hh pathway components by systematically testing gene function using RNAi-based technology.

刺猬（HH）蛋白质家族在许多人中协调细胞的生长和分化。 通过在产生HH产生细胞周围的细胞中引起分级反应的发育环境。异常 HH途径活动与面部和前脑的发育畸形有关，并且普遍存在 前列腺癌和基底细胞癌（BCC）等癌症。了解HH信号的方式 因此，从细胞外环境传达到细胞内效应子是成功的 与HH信号相关的这些疾病的预防，检测和治疗。这个建议 重点关注允许血浆中分泌HH蛋白的细胞识别的机制 膜以及随后在整个膜上的信号转导，以诱导特定的细胞内 回答。 HH与十二个跨膜蛋白（PTC）的直接相互作用是需要的 放弃PTC介导的抑制在靶标中七个跨膜蛋白平滑（SMO）的抑制 细胞。这种相互作用是由硫酸乙酰肝素蛋白聚糖和cg9211，a促进的蛋白质（DIP）促进的 免疫球蛋白（IG）受体超家族的成员。激活的SMO在跨整个 膜在涉及招募其细胞质尾部的过程中 转录因子Cubitus Intruptus（CI）。浸入CG9211的机制以及这些机制 细胞质蛋白 - 蛋白质相互作用有助于HH途径反应。 为了研究细胞膜HH信号的接收和传播，我们有 开发的新型工具，包括基于培养细胞的途径响应能力的培养细胞测定法， 分离和检测HH信号传导复合物以及抑制特定基因的双链RNA库 RNA介导的干扰（RNAi）在果蝇和小鼠中的功能。通过将这些工具合并到 基于生化和遗传学的策略，我们建议：1）确定如何在 细胞膜，2）确定SMO如何转导HH信号到细胞质成分，3）识别 HH途径组件通过使用基于RNAi的技术系统测试基因功能。