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2015 ; 13
(4
): e1002129
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GSK3?-Dzip1-Rab8 cascade regulates ciliogenesis after mitosis
#MMPMID25860027
Zhang B
; Zhang T
; Wang G
; Wang G
; Chi W
; Jiang Q
; Zhang C
PLoS Biol
2015[Apr]; 13
(4
): e1002129
PMID25860027
show ga
The primary cilium, which disassembles before mitotic entry and reassembles after
mitosis, organizes many signal transduction pathways that are crucial for cell
life and individual development. However, how ciliogenesis is regulated during
the cell cycle remains largely unknown. Here we show that GSK3?, Dzip1, and Rab8
co-regulate ciliogenesis by promoting the assembly of the ciliary membrane after
mitosis. Immunofluorescence and super-resolution microscopy showed that Dzip1 was
localized to the periciliary diffusion barrier and enriched at the mother
centriole. Knockdown of Dzip1 by short hairpin RNAs led to failed ciliary
localization of Rab8, and Rab8 accumulation at the basal body. Dzip1
preferentially bound to Rab8GDP and promoted its dissociation from its inhibitor
GDI2 at the pericentriolar region, as demonstrated by sucrose gradient
centrifugation of purified basal bodies, immunoprecipitation, and
acceptor-bleaching fluorescence resonance energy transfer assays. By means of in
vitro phosphorylation, in vivo gel shift, phospho-peptide identification by mass
spectrometry, and GST pulldown assays, we demonstrated that Dzip1 was
phosphorylated by GSK3? at S520 in G0 phase, which increased its binding to GDI2
to promote the release of Rab8GDP at the cilium base. Moreover, ciliogenesis was
inhibited by overexpression of the GSK3?-nonphosphorylatable Dzip1 mutant or by
disabling of GSK3? by specific inhibitors or knockout of GSK3? in cells.
Collectively, our data reveal a unique cascade consisting of GSK3?, Dzip1, and
Rab8 that regulates ciliogenesis after mitosis.