A Programmable DNA Origami Platform to Organize SNAREs for Membrane Fusion
Year: 2016
Type: article
Abstract: Soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) complexes are the core molecular machinery of membrane fusion, a fundamental process that drives inter- and intracellular communication and trafficking. One of the questions that remains controversial has been whether and how SNAREs cooperate. Here we show the use of self-assembled DNA-nanostructure rings to template uniform-sized small unilamellar vesicles containing predetermined maximal number of externally facing SNAREs to study the membrane-fusion process. We also incorporated lipid-conjugated complementary ssDNA as tethers into vesicle and target membranes, which enabled bypass of the rate-limiting docking step of fusion reactions and allowed direct observation of individual membrane-fusion events at SNARE densities as low as one pair per vesicle. With this platform, we confirmed at the single event level that, after docking of the templated-SUVs to supported lipid bilayers (SBL), one to two pairs of SNAREs are sufficient to drive fast lipid mixing. Modularity and programmability of this platform makes it readily amenable to studying more complicated systems where auxiliary proteins are involved.
DOI: 10.1021/jacs.5b13107
Authors: Weiming Xu, Bhavik Nathwani, Chenxiang Lin, Jing Wang, Erdem Karatekin, Frederic Pincet, William Shih, James E. Rothman
Institutions: Centre National de la Recherche Scientifique, Harvard University, Yale University, Neurophotonics Laboratory, Dana-Farber Cancer Institute, Laboratoire de Physique de l'Ecole Normale Supérieure, Université Paris Diderot, Université Paris Descartes, Université Pierre-et-Marie-Curie, Wyss Institute for Biologically Inspired Engineering
Language: English
Subfield: Cell Biology
Domain: Life Sciences
Open Access status: green
Funders: Yale University, Agence Nationale de la Recherche, National Institutes of Health, National Institute of General Medical Sciences, National Institute of Diabetes and Digestive and Kidney Diseases, National Center for Chronic Disease Prevention and Health Promotion, Division of Computing and Communication Foundations, Army Research Office
Awards: G1958771101, Generating nuclear pore complex mimics with DNA origami, Cell-free membrane remodeling guided by DNA nano-templates, Nucleation and dynamics of exocytotic fusion pores, Regulation of Membrane Fusion in Exocytosis, G5599132621, G5932455808, G6413502617, NMR Structure Determination of Membrane Proteins Enabled by DNA Nanotubes, G7828206644, Mechanisms of Intracellular Membrane Fusion
OpenAlex ID: W2290620539