Genetic Analysis of Legionella Phagosome Trafficking

军团菌吞噬体贩运的遗传分析

基本信息

批准号：
8721623
负责人：
Craig R. Roy
金额：
$ 8.82万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1997
资助国家：
美国
起止时间：
1997-07-01 至 2017-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8721623
关键词：
Abbreviations Bacteria Bacterial Model Bacterial Proteins Binding Biochemical Biogenesis Biological Assay Cell membrane Cell physiology Cells Cellular biology Complex Cyclic AMP Data Endoplasmic Reticulum Engineering Ensure Eukaryotic Cell Fluorescence Recovery After Photobleaching Funding Future GTPase-Activating Proteins Glutamate-ammonia-ligase adenylyltransferase Goals Growth Guanine Nucleotide Exchange Factors Guanosine Triphosphate Phosphohydrolases Health Human In Vitro Infection Integration Host Factors Legionella Legionella pneumophila Legionnaires&apos Disease Liquid Chromatography Lysosomes Mammalian Cell Mass Spectrum Analysis Measures Membrane Microbe Microscopy Molecular Nuclear Organelles Pathogenesis Pathway interactions Phagocytes Phagosomes Phosphoric Monoester Hydrolases Phosphorylcholine Phosphotransferases Play Pneumonia Post-Translational Protein Processing Process Protein Secretion Proteins Research Research Project Grants Role SNAP receptor System Transmembrane Transport Transport Vesicles Vacuole Vesicle combat genetic analysis maltose-binding protein novel pathogen phosphatidylinositol 4-phosphate protein function rab GTP-Binding Proteins research study soluble NSF attachment protein systems research tandem mass spectrometry trafficking

项目摘要

DESCRIPTION (provided by applicant): Legionella pneumophila is the causative agent of a severe pneumonia called Legionnaires' disease. The ability of Legionella to replicate inside of phagocytic cells is central to host pathogenesis and requires a specialized secretion system called Dot/Icm. The focus of this project has been to determine how the Dot/Icm system enables Legionella to create an intracellular vacuole that supports replication. Towards this end, we have demonstrated that this process involves the subversion of host vesicles derived from the endoplasmic reticulum, which are used by Legionella to remodel the plasma membrane-derived organelle it occupies initially into a vacuole that resembles the endoplasmic reticulum. Proteins delivered into host cells by the Dot/Icm system promote this membrane transport pathway. Over the past funding period we have identified multiple bacterial proteins that target the host membrane transport regulator Rab1. To understand how these proteins control membrane transport and promote biogenesis of a vacuole that permits Legionella intracellular growth, we will investigate the function of these proteins during infection of host cells by Legionella. Specifically, we will determine how the biochemical activities of these proteins are regulated spatially and temporally during infection to coordinate the cycling of Rab1 on vacuoles containing Legionella, determine how Rab1 activation on the vacuole containing Legionella can promote the recruitment and fusion of endoplasmic reticulum-derived vesicles, and determine the role host proteins that regulate phosphatidylinositol 4-phosphate play in vacuole maturation.

描述（由申请人提供）：肺炎军团菌是一种严重的肺炎的病因，称为军团疾病。军团菌在吞噬细胞内复制的能力对于宿主发病是至关重要的，需要一个称为DOT/ICM的专门分泌系统。该项目的重点是确定点/ICM系统如何使军团菌能够创建支持复制的细胞内液泡。为此，我们已经证明了这一过程涉及源自内质网的宿主囊泡，而Legionella则将其用于重塑质膜衍生的细胞器，其最初占据的真空吸尘器类似于内质网。 DOT/ICM系统输送到宿主细胞中的蛋白质促进了该膜转运途径。在过去的资金期间，我们已经确定了靶向宿主膜转运器RAB1的多种细菌蛋白。为了了解这些蛋白质如何控制膜转运并促进允许军团内生长的液泡的生物发生，我们将研究这些蛋白质在通过军团菌感染宿主细胞期间这些蛋白质的功能。 Specifically, we will determine how the biochemical activities of these proteins are regulated spatially and temporally during infection to coordinate the cycling of Rab1 on vacuoles containing Legionella, determine how Rab1 activation on the vacuole containing Legionella can promote the recruitment and fusion of endoplasmic reticulum-derived vesicles, and determine the role host proteins that regulate phosphatidylinositol 4-phosphate玩液泡成熟。