title: Early steps in primary cilium assembly require EHD1/EHD3-dependent ciliary vesicle formation.
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.
authors: NCI-Frederick National Laboratory, Laboratory of Cellular and Developmental Signaling, Frederick, MD 21702, USA.
authors: Quanlong
authors: Q
authors: Lu
authors: NCI-Frederick National Laboratory, Laboratory of Cellular and Developmental Signaling, Frederick, MD 21702, USA.
authors: Christine
authors: C
authors: Insinna
authors: Eunice Kennedy Shriver National Institute of Child Health and Human Development, NIH, Bethesda, MD.
authors: Carolyn
authors: C
authors: Ott
authors: NCI-Frederick National Laboratory, Laboratory of Cellular and Developmental Signaling, Frederick, MD 21702, USA.
authors: Jimmy
authors: J
authors: Stauffer
authors: CEDOC, Faculdade de Ciências Médicas, Universidade Nova de Lisboa, 1169-056 Lisboa, Portugal.
authors: Petra A
authors: PA
authors: Pintado
authors: Department of Biochemistry and Molecular Biology and the Fred and Pamela Buffett Cancer Center, University of Nebraska Medical Center, Omaha, Nebraska 68198, USA.
authors: Juliati
authors: J
authors: Rahajeng
authors: ATRF, NCI-Frederick National Laboratory, Frederick, MD.
authors: Ulrich
authors: U
authors: Baxa
authors: NCI-Frederick National Laboratory, Laboratory of Cellular and Developmental Signaling, Frederick, MD 21702, USA.
authors: Vijay
authors: V
authors: Walia
authors: NCI-Frederick National Laboratory, Laboratory of Cellular and Developmental Signaling, Frederick, MD 21702, USA.
authors: Adrian
authors: A
authors: Cuenca
authors: NCI-Frederick National Laboratory, Laboratory of Cellular and Developmental Signaling, Frederick, MD 21702, USA.
authors: Yoo-Seok
authors: YS
authors: Hwang
authors: NCI-Frederick National Laboratory, Laboratory of Cellular and Developmental Signaling, Frederick, MD 21702, USA.
authors: Ira O
authors: IO
authors: Daar
authors: ATRF, NCI-Frederick National Laboratory, Frederick, MD.
authors: Susana
authors: S
authors: Lopes
authors: Eunice Kennedy Shriver National Institute of Child Health and Human Development, NIH, Bethesda, MD.
authors: Jennifer
authors: J
authors: Lippincott-Schwartz
authors: Baxter Laboratory for Stem Cell Biology, Department of Microbiology and Immunology, Stanford University School of Medicine, Stanford, CA 94305.
authors: Peter K
authors: PK
authors: Jackson
authors: Department of Biochemistry and Molecular Biology and the Fred and Pamela Buffett Cancer Center, University of Nebraska Medical Center, Omaha, Nebraska 68198, USA.
authors: Steve
authors: S
authors: Caplan
authors: NCI-Frederick National Laboratory, Laboratory of Cellular and Developmental Signaling, Frederick, MD 21702, USA.
authors: Christopher J
authors: CJ
authors: Westlake
mesh_terms: D000818:Animals; D054468:Axoneme / Q000378:metabolism / Q000648:ultrastructure; D002352:Carrier Proteins / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D002460:Cell Line; D002502:Centrioles / Q000378:metabolism / Q000648:ultrastructure; D002923:Cilia / Q000378:metabolism* / Q000648:ultrastructure; D004625:Embryo, Nonmammalian; D004847:Epithelial Cells / Q000378:metabolism* / Q000648:ultrastructure; D018507:Gene Expression Regulation, Developmental; D006801:Humans; D007685:Kidney Tubules, Collecting / Q000378:metabolism / Q000648:ultrastructure; D051379:Mice; D009024:Morphogenesis / Q000235:genetics; D050766:Qb-SNARE Proteins / Q000235:genetics / Q000378:metabolism; D050767:Qc-SNARE Proteins / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D055213:Retinal Pigment Epithelium / Q000378:metabolism / Q000648:ultrastructure; D015398:Signal Transduction; D022161:Transport Vesicles / Q000378:metabolism* / Q000648:ultrastructure; D033921:Vesicular Transport Proteins / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D015027:Zebrafish; D020691:rab GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism; D000099193:rab11 GTP-Binding Proteins
chemical_list: D002352:Carrier Proteins; D050766:Qb-SNARE Proteins; D050767:Qc-SNARE Proteins; D034741:RNA, Small Interfering; D033921:Vesicular Transport Proteins; D020691:rab GTP-Binding Proteins; D000099193:rab11 GTP-Binding Proteins; C485640:Ehd1 protein, mouse; C470993:Ehd3 protein, mouse; C514104:Ift20 protein, mouse; C521976:Rab8 protein, mouse; C486621:Snap29 protein, mouse