Regulation of Germ Cell Fate During Embryogenesis

胚胎发生过程中生殖细胞命运的调节

基本信息

批准号：
7983744
负责人：
GERALDINE Catherine Joelle SEYDOUX
金额：
$ 27.22万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1999
资助国家：
美国
起止时间：
1999-08-18 至 2015-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7983744
关键词：
Anterior Architecture Automobile Driving Binding Biochemical Biochemical Genetics Biological Models Caenorhabditis elegans Cell Polarity Cell division Cells Complement Complex Cues Cytoplasm Cytoplasmic Granules Data Development Developmental Biology Diffuse Diffusion Embryo Embryonic Development Epithelial Cells Fluorescence Genes Genetic Techniques Germ Cells Goals In Vitro Lead Life Link Lymphocyte Microscopy Mitosis Modeling Neurons Phosphoric Monoester Hydrolases Phosphorylation Phosphorylation Site Phosphotransferases Proteins RNA-Binding Proteins Regulation Role Somatic Cell Specificity Spectrum Analysis System Testing Time Transcription Repressor/Corepressor cell type design human JTB protein human RBM5 protein in vivo mutant neuronal cell body polarized cell prevent protein aggregate protein complex public health relevance tumor uncontrolled cell growth zygote

项目摘要

DESCRIPTION (provided by applicant): The long-term goal of this project is to characterize the mechanisms that distinguish germ cells from somatic cells, a fundamental problem in developmental biology. In C. elegans, soma- germline asymmetries are established in the zygote before the first division, through the asymmetric partitioning of proteins and protein/RNA complexes in the cytoplasm. Cytoplasmic partitioning is regulated by conserved polarity regulators (PAR proteins), which localize asymmetrically in the zygote cortex. The goal of this proposal is to uncover how PAR activity at the cortex regulates polarity in the cytoplasm. Specific Aim I (SA1) will focus on the mechanisms that segregate the RNA-binding protein MEX-5 to the anterior cytoplasm. Preliminary data suggest that MEX-5 asymmetry depends on phosphorylation by PAR-1, a kinase on the posterior cortex, which increases MEX-5 diffusion locally in the posterior cytoplasm. SA2 will focus on the germline determinant PIE-1, which segregate to the posterior, opposite MEX-5 and in a steeper gradient. We will investigate how PIE-1 integrates both cortical and cytoplasmic cues to form a distinct gradient. Finally, SA3 will focus on the mechanisms that localize P granules to the posterior. P granules are evolutionarily conserved RNA-protein complexes specific to the germline. We have discovered a new P granule component PPTR-1 for P granule integrity during mitosis; our initial findings suggest that the germline specificity of P granules depends on regulated assembly and does not require cytoplasmic partitioning. We will use a combination of biochemical, genetic, and live microscopy approaches to test each of these hypotheses, and identify the genes and biochemical interactions involved. Unlike other well-studied embryonic polarity models, our system does not rely on pre-localized RNAs to generate asymmetry, and thus gives us the unique opportunity to explore the mechanisms that directly segregate proteins in the cytoplasm. PUBLIC HEALTH RELEVANCE: Cell polarity is essential to generate cell diversity during development, to prevent uncontrolled cell growth, and for the every-day functioning of many polarized cell types (epithelial cells, neurons, and lymphocytes). By taking advantage of a simple model system, our studies will illuminate the mechanisms that build and maintain the architecture of many cell types and prevent tumor formation.

描述（由申请人提供）：该项目的长期目标是表征将生殖细胞与体细胞区分开的机制，这是发育生物学的基本问题。在秀丽隐杆线虫中，通过蛋白质和细胞质中蛋白质/RNA复合物的不对称分配，在第一分裂之前的合子中建立了soma-生殖线不对称。细胞质分配受保守的极性调节剂（PAR蛋白）的调节，该蛋白在合子皮质中不对称地定位。该建议的目的是发现皮质的PAR活性如何调节细胞质的极性。具体的目标I（SA1）将集中在将RNA结合蛋白MEX-5分离为前细胞质的机制上。初步数据表明，MEX-5不对称依赖于PAR-1的磷酸化，PAR-1是后皮质上的激酶，该激酶在后细胞质中局部增加了MEX-5扩散。 SA2将专注于种系决定性pie-1，该pie-pie-1隔离到后5，在MEX-5对面，并且梯度较高。我们将研究PIE-1如何整合皮质和细胞质提示以形成独特的梯度。最后，SA3将集中在将P颗粒定位在后部的机制上。 P颗粒是特定于种系的进化保守的RNA-蛋白质复合物。我们发现了一个新的P颗粒成分PPTR-1，以用于有丝分裂过程中P颗粒完整性。我们的最初发现表明，P颗粒的种系特异性取决于调节的组装，不需要细胞质分配。我们将结合生化，遗传和实时显微镜方法来检验这些假设中的每一个，并确定所涉及的基因和生化相互作用。与其他良好的胚胎极性模型不同，我们的系统不依赖于预先定位的RNA来产生不对称性，因此为我们提供了独特的机会来探索直接隔离细胞质中蛋白质的机制。公共卫生相关性：细胞极性对于在发育过程中产生细胞多样性，预防不受控制的细胞生长以及许多极化细胞类型（上皮细胞，神经元和淋巴细胞）的每日功能至关重要。通过利用简单的模型系统，我们的研究将阐明构建和维护许多细胞类型的结构并防止肿瘤形成的机制。