Intercellular Signaling and Endosome to Golgi Transport in Multicellular Animals

多细胞动物的细胞间信号传导和内体到高尔基体的运输

基本信息

批准号：
8996179
负责人：
Barth Demian Grant
金额：
$ 29.45万
依托单位：
RUTGERS, THE STATE UNIV OF N.J.
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-02-01 至 2018-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8996179
关键词：
AMPA Receptors Actins Alzheimer&apos s Disease Animals Apoptotic Binding Caenorhabditis elegans Cell membrane Cell physiology Cells Centers for Disease Control and Prevention (U.S.)Clathrin Collaborations Complex Copper Data Development Disease Drosophila genus Endosomes Epithelial Event Feedback Generations Genetic Screening Goals Golgi Apparatus Guanosine Triphosphate Phosphohydrolases Health Human IGF Type 2 Receptor Immunity Ligands Link Lysosomal Storage Diseases Lysosomes Malignant Neoplasms Mammalian Cell Mediating Membrane Menkes Kinky Hair Syndrome Metabolism Microscopic Molecular Molecular Chaperones Nematoda Nervous system structure Organism Parkinson Disease Pathogenicity Pathway interactions Phylogenetic Analysis Process Proteins RNA interference screen Receptor Signaling Recycling Regulation Research Retrieval Ricin Role SH3 Domains Signal Pathway Signal Transduction Sorting - Cell Movement System Tertiary Protein Structure Testing Toxin Transducers Transforming Growth Factor beta WAVE protein Work Yeasts combat disorder control genetic regulatory protein genome-wide improved insight intestinal epithelium mutant novel protein complex protein function receptor retrograde transport therapeutic target trafficking

项目摘要

DESCRIPTION (provided by applicant): Our long-term goal in this proposal is to identify and understand the cellular components that transport internalized endocytic cargo from endosomes to the Golgi apparatus (retrograde recycling), and to understand how this process regulates, and is integrated within, intercellular signaling pathways. Retrograde recycling has recently been linked to multiple metazoan-specific processes such as the generation of Wnt signaling gradients, AMPA-receptor signaling in the nervous system, clearance of dead apoptotic cells, and regulation of epithelial polarity. Wnt signaling requires endosome-to-Golgi retrieval of the Wnt ligand chaperone MIG-14/Wntless, and the molecular details of MIG-14/Wntless recycling have proven to be highly conserved in C. elegans, Drosophila, and mammalian cells. Without retrograde recycling, transmembrane cargo proteins such as MIG-14/Wntless aberrantly enter the lysosome and are degraded, thus depleting the functional pool of such proteins from the cell. To gain novel insight into how retrograde recycling functions, and how it influences intercellular signaling mechanisms in multicellular animals, we have pioneered analysis of the polarized intestinal epithelium of C. elegans. In novel preliminary work we discovered that TGFβ-signaling is impaired in retrograde recycling mutants. We plan to further test the hypothesis that TGFβ-receptors are retrograde cargo, and test phylogenetic conservation of this mechanism in Drosophila and mammalian cells. In addition, our team has discovered previously unsuspected requirements for endosome regulatory proteins beyond the well-studied retromer complex. These novel regulators include the J-domain co-chaperone RME-8. We plan to test the hypothesis that RME-8 regulates the disassembly of the endosomal flat clathrin lattice, mediating a feedback loop between retromer and the ESCRT-driven degradation machinery that is present on the same endosomes. In addition we have identified an entire additional protein complex involved in retrograde recycling of MIG-14/Wntless. This protein complex appears to be associated with the recycling endosome, and contains modules for membrane binding, membrane bending, and actin nucleation. We plan to determine how this protein complex influences MIG-14/Wntless recycling, test its ability to influence other retrograde cargo including TGFβ-receptors, and test for conservation of function in mammalian cells. We expect that our findings will provide unique insight into how trafficking mechanisms influence signaling pathways, and will improve our ability to treat diverse diseases that arise from misregulation of receptor signaling strength, such as in cancers, and other diseases associated with misregulated retrograde recycling including Alzheimer's and Parkinson's diseases.

描述（由应用程序提供）：我们在此提案中的长期目标是识别和理解将内部内吞货物从内体传递到高尔基体设备（逆行回收）的细胞成分，并了解该过程如何调节并在内部集成细胞间信号通路中。逆行回收最近与多个后生动物特异性过程有关，例如神经系统中的Wnt信号传导梯度的产生，AMPA受体信号传导，死亡凋亡细胞的清除以及上皮极性的调节。 Wnt信号需要Wnt配体伴侣MIG-14/Wntless的内体至高尔基体检索，并且已证明Mig-14/Wnt无wnt无循环的分子细节已被证明是高度保守的秀丽隐杆线虫，果蝇，果蝇和哺乳动物细胞。在不逆行回收的情况下，跨膜货物蛋白（例如Mig-14/Wntless）异常进入溶酶体并降解，从而从细胞中耗尽了此类蛋白质的功能池。为了获得有关逆行回收功能以及它如何影响多细胞动物的细胞间信号传导机制的新洞察力，我们对秀丽隐杆线虫的偏振肠上皮进行了策划分析。在新的初步工作中，我们发现TGFβ信号在逆行回收突变体中受到损害。我们计划进一步检验以下假设：TGFβ受体是逆行货物，并在果蝇和哺乳动物细胞中测试该机制的系统发育保护。此外，我们的团队还发现了以前未经证实的对内体调节蛋白的要求，而不是研究良好的逆转录综合体。这些新颖的调节剂包括J-Domain共同伴侣RME-8。我们计划测试RME-8调节内体扁平网状晶格的拆卸的假设，从而介导了逆转录器和ESCRT驱动的降解机械之间的反馈回路，该反馈存在于同一内体上。此外，我们已经确定了参与MIG-14/Wntless逆行回收的整个额外蛋白质复合物。该蛋白质复合物似乎与回收内体有关，并包含用于膜结合，膜弯曲和肌动蛋白成核的模块。我们计划确定这种蛋白质复合物如何影响MIG-14/Wnt无回收，测试其影响其他逆行货物在内的逆行货物的能力，并测试哺乳动物细胞中功能的保护。我们预计我们的发现将提供有关运输机制如何影响信号通路的独特见解，并将提高我们治疗因受体信号强度不正常（例如在癌症中）以及其他与逆行逆行逆转的疾病（包括阿尔茨海默氏症和帕克森的疾病）相关的疾病所引起的潜水疾病的能力。