Abstract
Membrane association with mother centriole (M-centriole) distal appendages is critical for ciliogenesis initiation. How the Rab GTPase Rab11-Rab8 cascade functions in early ciliary membrane assembly is unknown. Here, we show that the membrane shaping proteins EHD1 and EHD3, in association with the Rab11-Rab8 cascade, function in early ciliogenesis. EHD1 and EHD3 localize to preciliary membranes and the ciliary pocket. EHD-dependent membrane tubulation is essential for ciliary vesicle formation from smaller distal appendage vesicles (DAVs). Importantly, this step functions in M-centriole to basal body transformation and recruitment of transition zone proteins and IFT20. SNAP29, a SNARE membrane fusion regulator and EHD1-binding protein, is also required for DAV-mediated ciliary vesicle assembly. Interestingly, only after ciliary vesicle assembly is Rab8 activated for ciliary growth. Our studies uncover molecular mechanisms informing a previously uncharacterized ciliogenesis step, whereby EHD1 and EHD3 reorganize the M-centriole and associated DAVs before coordinated ciliary membrane and axoneme growth.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, N.I.H., Intramural
MeSH terms
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Animals
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Axoneme / metabolism
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Axoneme / ultrastructure
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Carrier Proteins / antagonists & inhibitors
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Carrier Proteins / genetics*
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Carrier Proteins / metabolism
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Cell Line
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Centrioles / metabolism
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Centrioles / ultrastructure
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Cilia / metabolism*
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Cilia / ultrastructure
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Embryo, Nonmammalian
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Epithelial Cells / metabolism*
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Epithelial Cells / ultrastructure
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Gene Expression Regulation, Developmental*
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Humans
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Kidney Tubules, Collecting / metabolism
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Kidney Tubules, Collecting / ultrastructure
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Mice
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Morphogenesis / genetics
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Qb-SNARE Proteins / genetics
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Qb-SNARE Proteins / metabolism
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Qc-SNARE Proteins / genetics
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Qc-SNARE Proteins / metabolism
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RNA, Small Interfering / genetics
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RNA, Small Interfering / metabolism
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Retinal Pigment Epithelium / metabolism
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Retinal Pigment Epithelium / ultrastructure
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Signal Transduction
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Transport Vesicles / metabolism*
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Transport Vesicles / ultrastructure
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Vesicular Transport Proteins / antagonists & inhibitors
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Vesicular Transport Proteins / genetics*
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Vesicular Transport Proteins / metabolism
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Zebrafish
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rab GTP-Binding Proteins / genetics
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rab GTP-Binding Proteins / metabolism
Substances
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Carrier Proteins
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Ehd1 protein, mouse
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Ehd3 protein, mouse
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Ift20 protein, mouse
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Qb-SNARE Proteins
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Qc-SNARE Proteins
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RNA, Small Interfering
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Rab8 protein, mouse
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Snap29 protein, mouse
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Vesicular Transport Proteins
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rab11 protein
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rab GTP-Binding Proteins