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10:14
Biochemical Assays for Analyzing Activities of ATP-dependent Chromatin Remodeling Enzymes
Published on: October 25, 2014
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ATP-dependent force generation and membrane scission by ESCRT-III and Vps4
Johannes Schöneberg1,2,3, Mark Remec Pavlin2,4, Shannon Yan1,2
1Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA 94720, USA.
Science (New York, N.Y.)
|December 22, 2018
Summary
Endosomal sorting complexes (ESCRTs) drive membrane scission. This study shows ATP-fueled ESCRT-III and Vps4 assemblies generate force to sever membranes, verifying key biological predictions.
Area of Science:
- Cell Biology
- Biochemistry
- Molecular Motors
Background:
- Endosomal sorting complexes required for transport (ESCRTs) are crucial for membrane remodeling.
- ESCRT machinery mediates membrane scission in diverse cellular processes like viral budding and cytokinesis.
Purpose of the Study:
- To investigate the mechanism of membrane scission by ESCRT-III and Vps4.
- To directly visualize and verify the forces generated during ESCRT-mediated scission.
Main Methods:
- Reconstitution of ESCRT-III subunits (Snf7, Vps24, Vps2) and Vps4 in giant vesicles.
- Creation of membrane nanotubes for observing scission dynamics.
- Photo-uncaging of ATP to trigger ESCRT-mediated force generation and scission.
Main Results:
- ATP-dependent force generation within nanotubes led to membrane scission.
- Vps4 catalytic activity and its coupling to ESCRT-III were essential for scission.
- Imaging revealed Snf7 and Vps4 puncta correlated with force, constriction, and preceded scission.
Conclusions:
- Directly demonstrates that ATP-hydrolyzing assemblies of ESCRT-III and Vps4 sever membranes.
- Verifies the long-standing model of ESCRT-mediated membrane scission through force generation.
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