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- Title
Transition from hemifusion to pore opening is rate limiting for vacuole membrane fusion.
- Authors
Reese, Christoph; Mayer, Andreas
- Abstract
Fusion pore opening and expansion are considered the most energy-demanding steps in viral fusion. Whether this also applies to soluble N-ethyl-maleimide sensitive fusion protein altachment protein receptor (SNARE)- and Rab-dependent fusion events has been unknown. We have addressed the problem by characterizing the effects of lysophosphatidylcholine (LPC) and other late-stage inhibitors on lipid mixing and pore opening during vacuole fusion. LPC inhibits fusion by inducing positive curvature in the bilayer and changing its biophysical properties. The LPC block reversibly prevented formation of the hemifusion intermediate that allows lipid, but not content, mixing. Transition from hemifusion to pore opening was sensitive to guanosine-5′-(γ-thio)triphosphate. It required the vacuolar adenosine triphosphatase V0 sector and coincided with its transformation. Pore opening was rate limiting for the reaction. As with viral fusion, opening the fusion pore may be the most energy-demanding step for intracellular, SNARE-dependent fusion reactions, suggesting that fundamental aspects of lipid mixing and pore opening are related for both systems.
- Subjects
VIRUSES; CELL receptors; PROTEINS; LIPIDS; ADENOSINE triphosphatase
- Publication
Journal of Cell Biology, 2005, Vol 171, Issue 6, p961
- ISSN
0021-9525
- Publication type
Article
- DOI
10.1083/jcb.200510018