EGF receptor mediated signaling in Drosophila

果蝇中 EGF 受体介导的信号传导

• 批准号: 9204840
• 负责人: Gertrud M. Schupbach
• 金额: $ 36.72万
• 依托单位: PRINCETON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-02-01 至 2019-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9204840
• 关键词: Actins; Affect; Anterior; Antineoplastic Agents; Award; Binding; Binding Proteins; Biological; Biological Assay; Biological Models; CRISPR/Cas technology; Cell Communication; Cell Differentiation process; Cells; ChIP-seq; Cytoskeleton; Dependence; Development; Dorsal; Down-Regulation; Drosophila genus; Embryo; Epidermal Growth Factor Receptor; Epithelial; Epithelium; Funding; Gene Expression Regulation; Generations; Genes; Genetic; Genetic Transcription; Goals; Human; Malignant Neoplasms; Malignant neoplasm of lung; Mediating; Meiosis; Molecular; Monitor; Mutate; Mutation; Neurons; Normal Cell; Oocytes; Oogenesis; Organism; Ovarian Follicle; Pathway interactions; Pattern; Peptide Initiation Factors; Play; Process; Production; Protein Tyrosine Kinase; Proteins; RNA; Receptor Activation; Receptor Gene; Regulation; Research; Research Project Grants; Ribosomes; Role; Signal Pathway; Signal Transduction; Signaling Molecule; System; Tissues; To specify; Transcript; Translations; Up-Regulation; Work; cell type; cellular microvillus; egg; eukaryotic initiation factor-5B; eukaryotic peptide initiation factor-1A; experimental study; fight against; in vivo; malignant breast neoplasm; mutant; notch protein; novel therapeutics; public health relevance; receptor; receptor-mediated signaling; response

DESCRIPTION (provided by applicant): Cell-cell communication plays an important role in the development of many tissues. This research project investigates the mechanisms that activate the Drosophila Epidermal Growth Factor receptor (Egfr) during oogenesis, and the cellular pathways that mediate the response to this receptor tyrosine kinase in the ovarian follicle cells. We have shown that the Drosophila Egfr is expressed in the follicle cells and receives a highly controlled signal from the germline encoded by the gene gurken (grk). Restricted activation of the Egfr by Grk initiates several different follicle cell responses and is required for axis formaton of the egg and embryo. We have also shown that grk expression in oogenesis is regulated by developmental inputs as well as a meiotic checkpoint. Our goal is to study the molecular regulation of Grk production in the germline and to analyze the patterning and differentiation processes that are activated in the follicle cells in response to receptor activation. Our specific aims are: 1) The regulation of grk translation: We will identify proteins that bind to the grk RNA and regulate its translation using CRISPR technology. We will determine how the meiotic checkpoint regulates translation of grk and focus on the interaction of Vasa protein with the translation initiation factor eIF1A in the checkpoint activated germline. We will perform ribosome footprinting assays on the grk RNA which will allow us to determine the direct effects on ribosome binding by the developmental regulatory factors and by the meiotic checkpoint. 2) Analysis of the follicle cell response to EGFR activation. We will analyze the cooperation of the Egfr pathway with the Notch and JAK/STAT pathway during posterior follicle cell differentiation. We have isolated mutations that uncover interactions of Egfr signaling with the Notch pathway and will investigate these interactions at a molecular level. One focus is the transcriptional regulator Hamlet that acts downstream of Notch in neuronal cells. Transcriptional profiling and ChIP-Seq experiments will provide a global analysis of the cooperation between the signaling pathways. We will also analyze the cell biological response to Egfr signaling in the posterior follicle cells with a focus on the production of the posterior polarizing signal and its dependence on the actin cytoskeleton and microvilli. Mutations in checkpoint genes, as well as unregulated activation of the human Egfr have been implicated in several forms of cancer. Our work will elucidate the regulatory mechanisms that control the activity of this receptor in an in vivo situation. It will also analyze the interactions between Egfr and other signaling pathways and provide a molecular understanding of the downstream effector pathways operating in the follicle cell epithelium, a model system for epithelial development and differentiation.

描述（由申请人提供）：细胞间通讯在许多组织的发育中发挥着重要作用。该研究项目研究了在卵子发生过程中激活果蝇表皮生长因子受体 (Egfr) 的机制，以及介导卵泡细胞中对该受体酪氨酸激酶反应的细胞途径。我们已经证明，果蝇 Egfr 在滤泡细胞中表达，并接收来自 gurken (grk) 基因编码的种系的高度控制信号。 Grk 对 Egfr 的有限激活会引发几种不同的卵泡细胞反应，并且是卵子和胚胎轴形成所必需的。我们还表明，卵子发生中的 grk 表达受到发育输入以及减数分裂检查点的调节。我们的目标是研究种系中 Grk 产生的分子调控，并分析毛囊细胞响应受体激活而激活的模式和分化过程。我们的具体 目标是： 1) grk翻译的调控：我们将鉴定与grk RNA结合的蛋白质，并利用CRISPR技术调控其翻译。我们将确定减数分裂检查点如何调节 grk 的翻译，并重点关注 Vasa 蛋白与检查点激活种系中翻译起始因子 eIF1A 的相互作用。我们将对 grk RNA 进行核糖体足迹分析，这将使我们能够确定发育调节因子和减数分裂检查点对核糖体结合的直接影响。 2)分析滤泡细胞对EGFR激活的反应。我们将分析 Egfr 通路与 Notch 和 JAK/STAT 通路在后滤泡细胞分化过程中的配合。我们分离出突变，揭示了 Egfr 信号传导与 Notch 通路的相互作用，并将在分子水平上研究这些相互作用。焦点之一是在神经元细胞中作用于 Notch 下游的转录调节因子 Hamlet。转录分析和 ChIP-Seq 实验将对信号通路之间的合作提供全面分析。我们还将分析后滤泡细胞对 Egfr 信号传导的细胞生物学反应，重点关注后偏振信号的产生及其依赖性 肌动蛋白细胞骨架和微绒毛。检查点基因的突变以及人类 Egfr 的不受控制的激活与多种癌症有关。我们的工作将阐明在体内情况下控制该受体活性的调节机制。它还将分析 Egfr 和其他信号通路之间的相互作用，并提供对滤泡细胞上皮（上皮发育和分化的模型系统）中运行的下游效应器通路的分子理解。