title: A Programmable DNA Origami Platform to Organize SNAREs for Membrane Fusion.
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.
authors.forename: Weiming
authors.initials: W
authors.lastname: Xu
authors.affiliation: Wyss Institute for Biologically Inspired Engineering and Biological Chemistry and Molecular Pharmacology, Harvard Medical School, and Department of Cancer Biology, Dana Farber Cancer Institute , Boston, Massachusetts 02115, United States.
authors.forename: Bhavik
authors.initials: B
authors.lastname: Nathwani
authors.affiliation: Wyss Institute for Biologically Inspired Engineering and Biological Chemistry and Molecular Pharmacology, Harvard Medical School, and Department of Cancer Biology, Dana Farber Cancer Institute , Boston, Massachusetts 02115, United States.
authors.forename: Chenxiang
authors.initials: C
authors.lastname: Lin
authors.forename: Jing
authors.initials: J
authors.lastname: Wang
authors.affiliation: Laboratoire de Neurophotonique, Université Paris Descartes, Faculté des Sciences Fondamentales et Biomédicales, Centre National de la Recherche Scientifique (CNRS) UMR8250, 45, rue des Saints Pères, 75270 Cedex 06 Paris, France.
authors.forename: Erdem
authors.initials: E
authors.lastname: Karatekin
authors.affiliation: Laboratoire de Physique Statistique, Ecole Normale Supérieure de Paris, Université Pierre et Marie Curie, Université Paris Diderot, Centre National de la Recherche Scientifique, UMR 8550, 24 rue Lhomond, 75005 Paris, France.
authors.forename: Frederic
authors.initials: F
authors.lastname: Pincet
authors.affiliation: Wyss Institute for Biologically Inspired Engineering and Biological Chemistry and Molecular Pharmacology, Harvard Medical School, and Department of Cancer Biology, Dana Farber Cancer Institute , Boston, Massachusetts 02115, United States.
authors.forename: William
authors.initials: W
authors.lastname: Shih
authors.forename: James E
authors.initials: JE
authors.lastname: Rothman
mesh_terms: D004247:DNA / Q000378:metabolism*; D004277:DNA, Single-Stranded / Q000737:chemistry; D008051:Lipid Bilayers / Q000737:chemistry; D008081:Liposomes / Q000737:chemistry; D008561:Membrane Fusion; D011485:Protein Binding; D050600:SNARE Proteins / Q000378:metabolism*; D033921:Vesicular Transport Proteins / Q000378:metabolism*
chemical_list: D004277:DNA, Single-Stranded; D008051:Lipid Bilayers; D008081:Liposomes; D050600:SNARE Proteins; D033921:Vesicular Transport Proteins; D004247:DNA