INCAGE · In vitro reconstitution and single cell analysis of the Shigella-septin cage.
„Хоризонт 2020“ — Действия „Мария Склодовска-Кюри“
- Период
- 2017-08-01 → 2019-07-31
- Финансиране от ЕС
- 183 455 €
- Участници
- 2
- Схема
- MSCA-IF-EF-ST
Линиите свързват координатора с партньорите.
Накратко на български
Септиновите „клетки“ в организма улавят бактериите Shigella, за да ги насочат към разграждане. Разбирането на този механизъм помага да се разбере как клетките се защитават от бактериални инфекции.
Кратко обяснение, генерирано от езиков модел по текста на CORDIS. Оригиналът е по-долу.
Резултати накратко
In vitro reconstitution and single cell analysis of the Shigella-septin cage.
Autophagy is a highly conserved degradative process that breaks down cytosolic material inside double-membrane vesicles (called autophagosomes) by fusion with lysosomes. Selective autophagy is an important host defense mechanism that recognizes intracellular bacterial pathogens, such as Shigella flexneri, for targeting to degradation. To avoid autophagy, S. flexneri can polymerize host cell actin and form an actin tail that permits dissemination and cell-to-cell spread. Septins, a poorly understood component of the cytoskeleton that interacts with actin filaments and cellular membranes, entrap actin-polymerizing Shigella in cage-like structures for targeting to autophagy. However, the molecular determinants that underpin septin cage assembly are mostly unknown, and, the fate of septin cage-entrapped bacterium (replication, persistence, death) remains to be fully defined. Hosted by the Mostowy lab my project entitled "In vitro reconstitution and single cell analysis of the Shigella-septin cage (INCAGE)", aims to study factors controlling septin cage assembly and bacterial metabolism inside septin cages. I am investigating septin cage biology using bottom-up (using purified septin complexes in vitro and cell-free extracts) and top-down (using epithelial cells and different types of cutting-edge microscopy) approaches. For my Marie Curie postdoctoral project, I propose that in depth investigation of the Shigella-septin cage can lead to new septin biology, and new roles for the cytoskeleton in cell-autonomous immunity. INCAGE is an excellent platform to illuminate new concepts on the cell biology of Shigella that are difficult or impossible to address using whole cells or animal models. Understanding how our cells respond to bacterial infections can help to combat infectious diseases and control the spread of antibiotic resistance.
Текст от CORDIS, на английски · Данни: CORDIS, © Европейски съюз
Цел на проекта
Autophagy, an intracellular degradation process, is an important defence mechanism against pathogenic bacteria, including the Gram-negative enteroinvasive human pathogen Shigella flexneri. Work has shown that septins, a component of the cytoskeleton that interacts with membranes, entrap cytosolic Shigella in cage-like structures for targeting to autophagy. However, the factors mediating septin cage formation are mostly unknown. The Mostowy lab has recently discovered that mitochondria are required for efficient septin cage assembly and that ~50% of entrapped Shigella are metabolically inactive. The precise fate of septin cage-entrapped bacterium (replication, persistence, death) remains to be defined. For my project, called INCAGE, I will reconstitute the septin cage in vitro and study host and bacterial factors that modulate septin assembly. I will purify septin complexes and reconstitute septin assemblies (filaments, rings, cages) in vitro, and investigate how auxiliary components (actin, mitochondria, endoplasmic reticulum) influence septin cage assembly. This will reveal how septins recognize bacteria and how they assemble into cage-like structures. In parallel, I will follow septin cage-entrapped Shigella at the level of the single cell to study the lethal action of septin caging and uncover mechanisms that enable bacterial survival. I will test if the subpopulation of septin cage entrapped bacteria that stay alive, can replicate or led to the formation of persister cells (transiently refractory to stress) able to cope with host imposed stress (antibiotics, autophagy). Completion of these objectives will identify factors that promote septin cage assembly and its lethal action against entrapped bacteria. In depth understanding of septin cage assembly and its antibacterial activity can reveal important information about septin biology and cell-autonomous immunity, and help to design novel approaches to counteract bacterial infections.
Оригинален текст от CORDIS (на английски).
Участници
- LONDON SCHOOL OF HYGIENE AND TROPICAL MEDICINE ROYAL CHARTER · LondonКоординаторОбединеното кралство
- IMPERIAL COLLEGE OF SCIENCE TECHNOLOGY AND MEDICINE · LondonОбединеното кралство
Връзки
Данни: CORDIS, © Европейски съюз