CEP290 is essential for the initiation of ciliary transition zone assembly
Fig 5
The N-terminus of CEP290 recruits DZIP1 to regulate TZ assembly.
(A) Localization of DZIP1 in spermatocytes in the indicated genetic background. DZIP1-GFP was completely lost in cep2901 mutants and cby;dila double mutants, but existed in other mutants, including cep290ΔC mutants. γ-Tubulin (red) was used to label the BBs. Bar, 1 μm. (B) Localization of DZIP1 in sensory neurons in the indicated genetic background. In cep2901 mutants, DZIP1-GFP was completely lost. In cep290ΔC mutants, DZIP1 was present, but the signal was severely reduced in sensory neurons. 21A6 staining indicates the ciliary base in sensory neurons. Uncropped images can be found in S1 Raw Image. Bars, 1 μm. (C) The C-terminus of CEP290 is required to limit the DZIP1 signal at the ring centriole in spermatids. Bar, 1 μm. (D) Overexpression of the CEP290 N-terminus (aa 1–650) ameliorated the localization defects of DZIP1 in cep2901 mutants. Bars, 1 μm. (E) Overexpression of the CEP290 N-terminus rescued the localization of MKS1 in spermatocytes and spermatids in cep2901 mutants. Bars, 1 μm. (F) Overexpression of CEP290 N-terminus restored the connection between the BBs and the membrane in cep2901 mutants. CellMask was used to label the membrane, and UNC-GFP indicates the BBs. Bar, 10 μm. BB, basal body; CEP290, centrosomal protein 290; DZIP1, DAZ interacting zinc finger protein 1; MKS1, Meckel syndrome type 1; TZ, transition zone.