title: Individual vesicle fusion events mediated by lipid-anchored DNA.
abstract: Membrane fusion consists of a complex rearrangement of lipids and proteins that results in the merger of two lipid bilayers. We have developed a model system that employs synthetic DNA-lipid conjugates as a surrogate for the membrane proteins involved in the biological fusion reaction. We previously showed that complementary DNA-lipids, inserted into small unilamellar vesicles, can mediate membrane fusion in bulk. Here, we use a model membrane architecture developed in our lab to directly observe single-vesicle fusion events using fluorescence microscopy. In this system, a planar tethered membrane patch serves as the target membrane for incoming vesicles. This allows us to quantify the kinetics and characteristics of individual fusion events from the perspective of the lipids or the DNA-lipids involved in the process. We find that the fusion pathways are heterogeneous, with an arrested hemi-fusion state predominating, and we quantitate the outcome and rate of fusion events to construct a mechanistic model of DNA-mediated vesicle fusion. The waiting times between docking and fusion are distributed exponentially, suggesting that fusion occurs in a single step. Our analysis indicates that when two lipid bilayers are brought into close proximity, fusion occurs spontaneously, with little or no dependence on the number of DNA hybrids formed.
authors.affiliation: Department of Chemistry, Stanford University, Stanford, California, USA.
authors.forename: Bettina
authors.initials: B
authors.lastname: van Lengerich
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Rawle
authors.forename: Poul Martin
authors.initials: PM
authors.lastname: Bendix
authors.forename: Steven G
authors.initials: SG
authors.lastname: Boxer
mesh_terms: D002784:Cholesterol / Q000737:chemistry; D004247:DNA / Q000737:chemistry / Q000378:metabolism*; D008051:Lipid Bilayers / Q000737:chemistry / Q000378:metabolism; D008561:Membrane Fusion; D010713:Phosphatidylcholines / Q000737:chemistry; D010714:Phosphatidylethanolamines / Q000737:chemistry; D053835:Unilamellar Liposomes / Q000737:chemistry / Q000378:metabolism*
chemical_list: C410262:1,2-dioleoyl-glycero-3-phosphatidyl ethanolamine; D008051:Lipid Bilayers; D010713:Phosphatidylcholines; D010714:Phosphatidylethanolamines; D053835:Unilamellar Liposomes; D004247:DNA; D002784:Cholesterol; C017251:1,2-oleoylphosphatidylcholine