Cryo-EM of the dynamin polymer assembled on lipid membrane

Membrane fission is a fundamental process in the regulation and remodeling of cell membranes. Dynamin, a large GTPase, mediates membrane fission by assembling around, constricting and cleaving the necks of budding vesicles(1). Here, we report a 3.75 Å resolution cryo-EM structure of the membrane-associated helical polymer of human dynamin-1 in the GMPPCP bound state. The structure defines the helical symmetry of the dynamin polymer and the positions of the oligomeric interfaces, which were validated by cell-based endocytosis assays. Compared to the lipid-free tetramer form(2), membrane-associated dynamin binds to the lipid bilayer with its pleckstrin homology domain (PHD) and self-assembles across the helical rungs via the GTPase domain(3). Notably, interaction with the membrane and helical assembly is accommodated by a severely bent bundle signaling element (BSE), which connects the GTPase domain with the rest of the protein. The BSE conformation is asymmetric across the inter-rung GTPase interface, and is unique compared to all known nucleotide-bound states of dynamin. The structure suggests that the BSE bends from forces generated from the GTPase dimer interaction that are transferred across the stalk to the PHD and lipid membrane. Mutations disrupting the BSE kink impaired endocytosis. We also report a 10.1 Å resolution cryo-EM map of a super-constricted dynamin polymer showing localized conformational changes at the BSE and GTPase domains induced by GTP hydrolysis that drive membrane constriction. Altogether, the results provide a structural basis for dynamin’s mechanism of action on lipid membrane.