MYOII-DRIVEN FISSION · Reconstitution of Myosin IIA-driven membrane fission in vitro
7РП — „Хора“ (Действия „Мария Кюри“)
- Период
- 2014-03-01 → 2016-02-29
- Финансиране от ЕС
- 194 047 €
- Участници
- 1
- Схема
- MC-IEF
Линиите свързват координатора с партньорите.
Накратко на български
Механизмът, чрез който протеинът миозин IIA отделя транспортни везикули от мембраната на Голги аппарата, се изучава чрез лабораторни модели. Разбирането на този процес помага да се разбере как клетките регулират вътрешния си транспорт, което е от съществено значение за тяхното оцеляване.
Кратко обяснение, генерирано от езиков модел по текста на CORDIS. Оригиналът е по-долу.
Резултати накратко
Reconstitution of Myosin IIA-driven membrane fission in vitro
Eukaryotic cells are subdivided in distinct membrane enclosed compartments that are connected by intracellular transport processes facilitated by vesicles. The temporal and spatial regulation of these transport processes is essential for the survival of cells. The transport vesicles are fissioned from the donor membrane after their formation. This events seems to be coordinated with the elongation of tubular precursors pulled by myosins and/or kinesins. Recent results show an unexpected function of nonmusle Myosin II A (nmMyoIIA) in fission of Rab6A transport carriers at the trans-Golgi network. The aim of this project is the development of an in vitro system to mimic the fission of vesicles from the trans-Golgi network. We have developed a system in which we can recruit Rab6A to Giant Unilamellar Vesicles (GUV) after prenylation of the protein in vitro. Several Rab6 effectors can be recruited to these GUVs in a Rab6:GTP dependent manner. Furthermore we have isolated Golgi membranes on which we were able to detect Rab6A, nmMyoIIA and actin and have pulled tubes from these membranes.
Текст от CORDIS, на английски · Данни: CORDIS, © Европейски съюз
Цел на проекта
Eukaryotic cells contain a variety of membrane-enclosed compartments that are connected by dynamic transport processes facilitated by vesicles. The temporal and spatial regulation of these transport processes is essential for cellular function. After initial formation transport vesicles are separated from their donor membrane and this fission event seems to be coordinated with the elongation of tubular precursors pulled by motor proteins. Recent results from the host laboratory demonstrated a direct interaction of Rab6 (a Golgi-associated RabGTPase) and nonmuscle Myosin II A (MyoIIA) and showed an unexpected function of MyoIIA in the fission of Rab6 transport carriers at the trans-Golgi network.The aim of this project is the development of an in vitro system to mimic the MyoIIA-driven fission process. In this system, we will pull membrane nanotubes from Giant Unilamellar Vesicles (GUVs) using optical tweezers and we will monitor tube stability in the presence of Rab6, MyoIIA, actin and ATP. Two hypotheses will be tested: either MyoIIA induces phase separation in membrane tubes by the formation of actomyosin patches, which then leads to fission; or MyoIIA contracts short actin filaments around the membrane tubes in a manner similar to its role during the formation of the cleavage furrow in cytokinesis. If we do not observe fission in this minimal system, we will add additional factors that have been implicated in fission at the trans-Golgi network.In the past years the cell biology host laboratory of Bruno Goud and the experimental physicists from Patricia Bassereau’s group have collaborated extensively to develop new interdisciplinary methods to investigate intracellular transport processes. As I have so far focused on biophysical measurements with Rab proteins in the absence of membranes, the development of the in vitro system in these laboratories will give me the possibility to get exposure to a variety of new techniques that will expand my current expertise.
Оригинален текст от CORDIS (на английски).
Участници
- INSTITUT CURIE · ParisКоординаторФранция
Връзки
Данни: CORDIS, © Европейски съюз