Molecular Determinants of TLR Trafficking

TLR 贩运的分子决定因素

基本信息

批准号：
9120303
负责人：
Gregory M Barton
金额：
$ 76.96万
依托单位：
SANFORD BURNHAM PREBYS MEDICAL DISCOVERY INSTITUTE
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-08-01 至 2018-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9120303
关键词：
Address Autoimmune Diseases Autoimmunity B-Lymphocytes Biochemical Biochemistry Biological Assay Carrier Proteins Cells Cellular biology Cleaved cell Complex DNA Data Dendritic Cells Development Disease Drug Targeting Early Endosome Endoplasmic Reticulum Endosomes Fractionation Genes Genetic Screening Golgi Apparatus Health Immune Immune response Indirect Immunofluorescence Infection Integral Membrane Protein Intestines Knock-in Mouse Knowledge Mediating Membrane Microscopy Modification Molecular Molecular Chaperones Multivesicular Body Onset of illness Pathway interactions Pattern recognition receptor Phagosomes Play Population Process Proteins Publishing RNA interference screen Recruitment Activity Role Signal Pathway Signal Transduction Sorting - Cell Movement Stromal Cells Surface System TLR5 gene TLR7 gene Tumor Necrosis Factor Receptor Ubiquitin Ubiquitination Vesicle base cell type cofactor drug development functional genomics genetic approach genome-wide hepatocyte growth factor-regulated tyrosine kinase substrate in vivo late endosome macrophage microbial pathogen receptor research study sensor trafficking ubiquitin ligase ubiquitin-protein ligase

项目摘要

DESCRIPTION (provided by applicant): TLR9 is an endosomally localized Pattern Recognition Receptor (PRR) that detects pathogen-associated DNA. TLR9 traffics from in the Endoplasmic Reticulum (ER) to the endolysosome, where it is cleaved and made competent for signaling activation. We have recently demonstrated that TLR9 is ubiquitinated and targeting of TLR9 to the early endosome is an ubiquitin-dependent process that is mediated by the ESCRT protein Hepatocyte Growth Factor Regulated Tyrosine Kinase Substrate (HRS). This compartmentalization is critical in mediating the signaling activation, as well as reducing the likelihood of triggering autoimmune disease. However, the molecular cofactors that enable TLR9 to transit from the ER to the endolysome are mostly uncharacterized. Here, we propose to identify the E3 ubiquitin ligase that is required for TLR9 ubiquitination using a high throughput genetic approach, as well as characterize the non-canonical role of ESCRT pathways components in TLR9 trafficking. In addition, we propose to validate 15 proteins identified by a genome-wide RNAi screen for TLR7/9 signaling for their potential roles in TLR7/9 endosomal trafficking. Finally, we propose to study the in vivo role of TLR9 ubiquitination and ESCRT function in TLR9 expressing cell subsets. Understanding the trafficking system of TLR9 is critical for the development of drugs for autoimmune diseases that results from promiscuous activation through endosomal TLR signaling.

描述（由申请人提供）：TLR9是检测与病原体相关DNA的内体局部局部模式识别受体（PRR）。 TLR9从内质网中（ER）流量到内溶液体，在那里裂解并使信号激活具有胜任。我们最近证明，TLR9是泛素化的，TLR9靶向早期内体是泛素依赖性的过程，是由ESCRT蛋白肝细胞生长因子调节的酪氨酸激酶底物（HRS）介导的。这种分区化对于介导信号激活以及降低触发自身免疫性疾病的可能性至关重要。但是，使TLR9从ER转到内溶溶裂体的分子辅助因子大多没有表征。在这里，我们建议使用高吞吐量遗传方法来识别TLR9泛素化所需的E3泛素连接酶，并表征TLR9运输中ESCRT途径成分的非典型作用。此外，我们建议通过全基因组RNAi筛选鉴定出15种蛋白质，以实现TLR7/9信号传导，以便其在TLR7/9内体运输中的潜在作用。最后，我们建议研究TLR9泛素化和ESCRT功能在TLR9表达细胞亚群中的体内作用。了解TLR9的运输系统对于通过内体TLR信号传导引起的自身免疫性疾病的药物开发至关重要。