10.5061/DRYAD.GN250
Adell, Manuel Alonso Y.
University of Innsbruck
Migliano, Simona M.
University of Innsbruck
Upadhyayula, Srigokul
Harvard Medical School
Bykov, Yury S.
European Molecular Biology Laboratory
Sprenger, Simon
University of Innsbruck
Pakdel, Mehrshad
Max Planck Institute of Biochemistry
University of Innsbruck
Vogel, Georg F.
University of Innsbruck
Jih, Gloria
Harvard Medical School
Skillern, Wesley
University of Innsbruck
Behrouzi, Reza
Harvard Medical School
Babst, Markus
University of Utah
Schmidt, Oliver
University of Innsbruck
Hess, Michael W.
University of Innsbruck
Briggs, John A.G.
European Molecular Biology Laboratory
Kirchhausen, Tomas
Harvard Medical School
Teis, David
University of Innsbruck
Briggs, John AG
European Molecular Biology Laboratory
Data from: Recruitment dynamics of ESCRT-III and Vps4 to endosomes and
implications for reverse membrane budding
Dryad
dataset
2017
Cell biology
S. cerevisiae
2017-10-30T18:08:12Z
2017-10-30T18:08:12Z
en
https://doi.org/10.7554/elife.31652
6810599916 bytes
1
CC0 1.0 Universal (CC0 1.0) Public Domain Dedication
The ESCRT machinery mediates reverse membrane scission. By quantitative
fluorescence lattice light-sheet microscopy, we have shown that ESCRT-III
subunits polymerize rapidly on yeast endosomes, together with the
recruitment of at least two Vps4 hexamers. During their 3-45 second
lifetimes, the ESCRT-III assemblies accumulated 75-200 Snf7 and 15-50
Vps24 molecules. Productive budding events required at least two
additional Vps4 hexamers. Membrane budding was associated with continuous,
stochastic exchange of Vps4 and ESCRT-III components, rather than steady
growth of fixed assemblies, and depended on Vps4 ATPase activity. An
all-or-none step led to final release of ESCRT-III and Vps4. Tomographic
electron microscopy demonstrated that acute disruption of Vps4 recruitment
stalled membrane budding. We propose a model in which multiple Vps4
hexamers (four or more) draw together several ESCRT-III filaments. This
process induces cargo crowding and inward membrane buckling, followed by
constriction of the nascent bud neck and ultimately ILV generation by
vesicle fission.
source fileNotes: 1. includes all detected and tracked object data from
deskewed volumes. 2. Analyzed data excluded objects at volume boundaries,
cut tracks and compound tracks. 3. Fitted amplitudes are converted to
single molecule values from the calibration curve generated for each power
condition before each experiment.Source file.matSource File notes.xlsx