Genetic Analysis of Legionella Phagosome Trafficking

军团菌吞噬体贩运的遗传分析

• 批准号: 10213614
• 负责人: Craig R. Roy
• 金额: $ 54.42万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-01 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10213614
• 关键词: Bacterial Proteins; Biochemical; Biogenesis; Cell membrane; Cells; Endoplasmic Reticulum; Eukaryotic Cell; Funding; Growth; Guanosine Triphosphate Phosphohydrolases; Immune system; In Vitro; Infection; Legionella; Legionella pneumophila; Legionnaires' Disease; Molecular; Organelles; Pathogenesis; Pathway interactions; Phagocytes; Phagosomes; Play; Pneumonia; Process; Proteins; Role; System; Transmembrane Transport; Vacuole; Vesicle; genetic analysis; novel; pathogen; phosphatidylinositol 4-phosphate; protein function; recruit; trafficking; vesicle transport

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.

嗜肺军团菌是一种称为军团病的严重肺炎的病原体。军团菌在吞噬细胞内复制的能力是宿主发病机制的核心，需要一个称为Dot/Icm的专门分泌系统。该项目的重点是确定Dot/Icm系统如何使军团菌产生支持复制的细胞内空泡。为此，我们已经证明，这一过程涉及宿主囊泡的颠覆来自内质网，这是由军团菌重塑质膜衍生的细胞器，它最初占据成一个类似于内质网的液泡。通过Dot/Icm系统递送到宿主细胞中的蛋白质促进该膜转运途径。在过去的资助期间，我们已经确定了多种靶向宿主膜转运调节因子Rab 1的细菌蛋白。为了了解这些蛋白质如何控制膜运输和促进生物发生的空泡，允许军团菌胞内生长，我们将调查这些蛋白质的功能，在军团菌感染宿主细胞。具体而言，我们将确定如何在感染过程中的空间和时间调节这些蛋白质的生化活性，以协调含有军团菌的空泡上Rab 1的循环，确定如何Rab 1激活含有军团菌的空泡可以促进内质网衍生的囊泡的招募和融合，并确定宿主蛋白调节磷脂酰肌醇4-磷酸在空泡成熟中发挥的作用。

项目成果

Legionella DotM structure reveals a role in effector recruiting to the Type 4B secretion system.

• DOI: 10.1038/s41467-017-02578-x
• 发表时间: 2018-02-06
• 期刊: Nature communications
• 影响因子: 16.6
• 作者: Meir A;Chetrit D;Liu L;Roy CR;Waksman G
• 通讯作者: Waksman G

DNA Delivery and Genomic Integration into Mammalian Target Cells through Type IV A and B Secretion Systems of Human Pathogens.

通过人类病原体的IV A和B分泌系统，DNA递送和基因组整合到哺乳动物靶细胞中。

• DOI: 10.3389/fmicb.2017.01503
• 发表时间: 2017
• 期刊: Frontiers in microbiology
• 影响因子: 5.2
• 作者: Guzmán-Herrador DL;Steiner S;Alperi A;González-Prieto C;Roy CR;Llosa M
• 通讯作者: Llosa M

Autophagy Evasion and Endoplasmic Reticulum Subversion: The Yin and Yang of Legionella Intracellular Infection.

自噬逃避和内质网颠覆：军团菌细胞内感染的阴阳。

• DOI: 10.1146/annurev-micro-102215-095557
• 发表时间: 2016
• 期刊: Annual review of microbiology
• 影响因子: 10.5
• 作者: Sherwood,RacquelKim;Roy,CraigR
• 通讯作者: Roy,CraigR

Proteins DotY and DotZ modulate the dynamics and localization of the type IVB coupling complex of Legionella pneumophila.

• DOI: 10.1111/mmi.14847
• 发表时间: 2022-03
• 期刊: MOLECULAR MICROBIOLOGY
• 影响因子: 3.6
• 作者: Mace, Kevin;Meir, Amit;Lukoyanova, Natalya;Liu, Luying;Chetrit, David;Hospenthal, Manuela K.;Roy, Craig R.;Waksman, Gabriel
• 通讯作者: Waksman, Gabriel

AMPylation is critical for Rab1 localization to vacuoles containing Legionella pneumophila.

• DOI: 10.1128/mbio.01035-13
• 发表时间: 2014-02-11
• 期刊: mBio
• 影响因子: 6.4
• 作者: Hardiman CA;Roy CR
• 通讯作者: Roy CR