Stepwise remodeling and subcompartment formation in individual vesicles by three ESCRT-III proteins
Yunuen Avalos-Padilla,
Vasil N. Georgiev,
Eleanor Ewins,
Tom Robinson,
Esther Orozco,
Reinhard Lipowsky,
Rumiana Dimova
Affiliations
Yunuen Avalos-Padilla
Max Planck Institute of Colloids and Interfaces, Science Park Golm, 14476 Potsdam, Germany; Nanomalaria Group, Institute for Bioengineering of Catalonia (IBEC), The Barcelona Institute of Science and Technology, Baldiri Reixac 10-12, ES-08028 Barcelona, Spain; Barcelona Institute for Global Health (ISGlobal, Hospital Clínic-Universitat de Barcelona), Rosselló 149-153, ES-08036 Barcelona, Spain
Vasil N. Georgiev
Max Planck Institute of Colloids and Interfaces, Science Park Golm, 14476 Potsdam, Germany
Eleanor Ewins
Max Planck Institute of Colloids and Interfaces, Science Park Golm, 14476 Potsdam, Germany
Tom Robinson
Max Planck Institute of Colloids and Interfaces, Science Park Golm, 14476 Potsdam, Germany
Esther Orozco
Departamento de Infectómica y Patogénesis Molecular, CINVESTAV IPN, 07360 Ciudad de México, México
Reinhard Lipowsky
Max Planck Institute of Colloids and Interfaces, Science Park Golm, 14476 Potsdam, Germany
Rumiana Dimova
Max Planck Institute of Colloids and Interfaces, Science Park Golm, 14476 Potsdam, Germany; Corresponding author
Summary: The endosomal sorting complex required for transport (ESCRT) is a multi-protein machinery involved in several membrane remodeling processes. Different approaches have been used to resolve how ESCRT proteins scission membranes. However, the underlying mechanisms generating membrane deformations are still a matter of debate. Here, giant unilamellar vesicles, microfluidic technology, and micropipette aspiration are combined to continuously follow the ESCRT-III-mediated membrane remodeling on the single-vesicle level for the first time. With this approach, we identify different mechanisms by which a minimal set of three ESCRT-III proteins from Entamoeba histolytica reshape the membrane. These proteins modulate the membrane stiffness and spontaneous curvature to regulate bud size and generate intraluminal vesicles even in the absence of ATP. We demonstrate that the bud stability depends on the protein concentration and membrane tension. The approaches introduced here should open the road to diverse applications in synthetic biology for establishing artificial cells with several membrane compartments.
