GTP hydrolysis promotes disassembly of the atlastin crossover dimer during ER fusion

Abstract

The GTPase atlastin mediates homotypic ER fusion through trans-crossover dimerization, but how dimerization is coupled to the GTPase cycle has remained unclear. Winsor et al. show that GTP binding causes crossover dimerization for fusion, whereas GTP hydrolysis promotes disassembly of the crossover dimer for subunit recycling.

Abstract

Membrane fusion of the ER is catalyzed when atlastin GTPases anchored in opposing membranes dimerize and undergo a crossed over conformational rearrangement that draws the bilayers together. Previous studies have suggested that GTP hydrolysis triggers crossover dimerization, thus directly driving fusion. In this study, we make the surprising observations that WT atlastin undergoes crossover dimerization before hydrolyzing GTP and that nucleotide hydrolysis and Pi release coincide more closely with dimer disassembly. These findings suggest that GTP binding, rather than its hydrolysis, triggers crossover dimerization for fusion. In support, a new hydrolysis-deficient atlastin variant undergoes rapid GTP-dependent crossover dimerization and catalyzes fusion at an initial rate similar to WT atlastin. However, the variant cannot sustain fusion activity over time, implying a defect in subunit recycling. We suggest that GTP binding induces an atlastin conformational change that favors crossover dimerization for fusion and that the input of energy from nucleotide hydrolysis promotes complex disassembly for subunit recycling.