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

• 批准号: 8608558
• 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
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-02-01 至 2017-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8608558
• 关键词: AMPA Receptors; Actins; Alzheimer' 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; PARD6A gene; Parkinson Disease; Pathogenicity; Pathway interactions; Phylogenetic Analysis; Process; Proteins; RNA Interference; Receptor Signaling; Recycling; Regulation; Research; Retrieval; Ricin; Role; SH3 Domains; Signal Pathway; Signal Transduction; Sorting - Cell Movement; System; Tertiary Protein Structure; Testing; Toxin; Transducers; WAVE protein; Work; Yeasts; combat; disorder control; genetic regulatory protein; genome-wide; improved; insight; intestinal epithelium; mutant; novel; protein complex; protein function; public health relevance; receptor; retrograde transport; therapeutic target; trafficking; AMPA Receptors; Actins; Alzheimer' 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; PARD6A gene; Parkinson Disease; Pathogenicity; Pathway interactions; Phylogenetic Analysis; Process; Proteins; RNA Interference; Receptor Signaling; Recycling; Regulation; Research; Retrieval; Ricin; Role; SH3 Domains; Signal Pathway; Signal Transduction; Sorting - Cell Movement; System; Tertiary Protein Structure; Testing; Toxin; Transducers; WAVE protein; Work; Yeasts; combat; disorder control; genetic regulatory protein; genome-wide; improved; insight; intestinal epithelium; mutant; novel; protein complex; protein function; public health relevance; receptor; retrograde transport; therapeutic target; trafficking

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 TGF?-signaling is impaired in retrograde recycling mutants. We plan to further test the hypothesis that TGF?-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 TGF?-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（由申请人提供）：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开创性的分析。在新的初步工作中，我们发现TGF？ - 信号在逆行回收突变体中受到损害。我们计划进一步测试TGF？受体是逆行货物的假设，并测试了果蝇和哺乳动物细胞中该机制的系统发育保守。此外，我们的团队已经发现了以前对内体调节蛋白超出良好的逆转录综合体以外的内体调节蛋白的要求。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（包括TGF？ - 受体受体）的能力，并测试哺乳动物细胞中功能的保存。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的不正调而引起的多种疾病的能力，例如在癌症中，以及其他与逆转不良的逆行相关的疾病，包括阿尔茨海默氏症和帕克森的疾病。