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GTP hydrolysis\u2010triggered conformational changes in ER\u2010anchored atlastins (ATLs) have been shown to be responsible for this process, which is facilitated by the amphipathic helix within its C\u2010terminal tail (AH\n                    <jats:sub>ATL\u2010C<\/jats:sub>\n                    ). However, how the multiple ATL molecules cooperate to complete membrane tethering and subsequent fusion is still not known. Here, we used a programmable DNA origami ring to generate uniformly sized proteoliposomes containing a controlled maximum number of\n                    <jats:italic>Drosophila<\/jats:italic>\n                    ATL (dmATL) molecules. Using this system, we demonstrated at the single\u2010event level that one pair of dmATL molecules is capable of mediating membrane tethering but not lipid mixing. To investigate the subsequent fusion event, we developed programmable DNA soccer\u2010ball frameworks as a template for the generation of tethered liposomes. With a combination of molecular dynamics simulations and DNA\u2010ring\u2010templated single\u2010vesicle fusion assays, we found that the AH\n                    <jats:sub>ATL\u2010C<\/jats:sub>\n                    of dmATL is sufficient to promote the merging of closely apposed membranes by enhancing bilayer fluctuations under specific lipid composition condition. Our results provide mechanistic insights into dmATL\u2010mediated homotypic membrane fusion, and our adaptable DNA nanostructures offer powerful tools for studying other biological processes requiring predetermined numbers of molecules or involving tethered membranes.\n                  <\/jats:p>","DOI":"10.1002\/smll.75007","type":"journal-article","created":{"date-parts":[[2026,8,3]],"date-time":"2026-08-03T09:57:40Z","timestamp":1785751060000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Organizing Large GTPase Atlastin\u2010Mediated Membrane Fusion by Programmable DNA Origami Platforms"],"prefix":"10.1002","volume":"22","author":[{"given":"Jingyao","family":"Li","sequence":"first","affiliation":[{"name":"State Key Laboratory of Medicinal Chemical Biology and College of Life Sciences Nankai University  Tianjin 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