VPS4 is a dynamic component of the centrosome that regulates centrosome localization of γ-tubulin, centriolar satellite stability and ciliogenesis

Scientific Reports
Carolyn OttNatalie Elia

Abstract

The hexameric AAA ATPase VPS4 facilitates ESCRT III filament disassembly on diverse intracellular membranes. ESCRT III components and VPS4 have been localized to the ciliary transition zone and spindle poles and reported to affect centrosome duplication and spindle pole stability. How the canonical ESCRT pathway could mediate these events is unclear. We studied the association of VPS4 with centrosomes and found that GFP-VPS4 was a dynamic component of both mother and daughter centrioles. A mutant, VPS4EQ, which can't hydrolyze ATP, was less dynamic and accumulated at centrosomes. Centrosome localization of the VPS4EQ mutant, caused reduced γ-tubulin levels at centrosomes and consequently decreased microtubule growth and altered centrosome positioning. In addition, preventing VPS4 ATP hydrolysis nearly eliminated centriolar satellites and paused ciliogensis after formation of the ciliary vesicle. Zebrafish embryos injected with GFP-VPS4EQ mRNA were less viable, exhibited developmental defects and had fewer cilia in Kupffer's vesicle. Surprisingly, ESCRT III proteins seldom localized to centrosomes and their depletion did not lead to these phenotypes. Our data support an ESCRT III-independent function for VPS4 at the centrosome a...Continue Reading

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Citations

Mar 13, 2020·PLoS Biology·Delphine GogendeauAnne-Marie Tassin
Jan 4, 2020·Journal of Cell Science·Suzanna L Prosser, Laurence Pelletier
Sep 19, 2018·Development·Lei Wang, Brian D Dynlacht
Jan 10, 2020·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Eunji JungJoon Kim
Feb 16, 2021·Frontiers in Cell and Developmental Biology·Leticia Labat-de-HozMiguel A Alonso

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Methods Mentioned

BETA
confocal microscopy
transfection
fluorescent recovery after photobleaching
electron microscopy
fluorescence microscopy
acetylation
transfections
PCR
Protein Assay
in-vitro transcription

Software Mentioned

SlideBook
Volocity
GraphPad Prism
Volocity6
ZEN
Image J
Airyscan
BLAST
Statistica Statistica

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