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FAP206 is a microtubule-docking adapter for ciliary radial spoke 2 and dynein c.

Vasudevan KK, Song K, Alford LM, Sale WS, Dymek EE, Smith EF, Hennessey T, Joachimiak E, Urbanska P, Wloga D, Dentler W, Nicastro D, Gaertig J - Mol. Biol. Cell (2014)

Bottom Line: A knockout of the conserved ciliary protein FAP206 in the ciliate Tetrahymena resulted in slow cell motility.Occasionally, RS2 assembled but lacked both the front prong of its microtubule base and dynein c, whose tail is attached to the front prong.Overexpressed GFP-FAP206 decorated nonciliary microtubules in vivo.

View Article: PubMed Central - PubMed

Affiliation: Department of Cellular Biology, University of Georgia, Athens, GA 30602.

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FAP206 localizes to the ciliary axoneme, and knockout of FAP206 results in cilia-related defects. (A) TIRF image of a live cell expressing FAP206-GFP under the native promoter. (B) A wild-type (negative control) cell (left) and a cell expressing FAP206-GFP under the native promoter (right) extracted with Triton X-100, fixed with paraformaldehyde, and imaged for GFP using a confocal microscope. (C) Culture growth rates for a wild-type CU428 and an FAP206-KO strain. Each data point represents an average for three independent experiments. (D) Paths of swimming wild-type and FAP206-KO cells recorded for 1 s. The average swim velocities were 170 μm/s for the wild type and 50 μm/s for FAP206-KO. (E) Immunofluorescence images of tubulin for wild-type and FAP206-KO cells. For each genotype, an interphase (left) and a dividing cell (right) are shown. (F) Classical TEM images of cilia cross-sections that are either circular (left) or compressed (right). The percentages represent fractions of either circular or compressed axonemes (n = 52 for wild type, n = 48 for FAP206-KO). Scale bars, 20 μm (A, B, and E), 1 mm (D), 0.2 μm (F).
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Figure 1: FAP206 localizes to the ciliary axoneme, and knockout of FAP206 results in cilia-related defects. (A) TIRF image of a live cell expressing FAP206-GFP under the native promoter. (B) A wild-type (negative control) cell (left) and a cell expressing FAP206-GFP under the native promoter (right) extracted with Triton X-100, fixed with paraformaldehyde, and imaged for GFP using a confocal microscope. (C) Culture growth rates for a wild-type CU428 and an FAP206-KO strain. Each data point represents an average for three independent experiments. (D) Paths of swimming wild-type and FAP206-KO cells recorded for 1 s. The average swim velocities were 170 μm/s for the wild type and 50 μm/s for FAP206-KO. (E) Immunofluorescence images of tubulin for wild-type and FAP206-KO cells. For each genotype, an interphase (left) and a dividing cell (right) are shown. (F) Classical TEM images of cilia cross-sections that are either circular (left) or compressed (right). The percentages represent fractions of either circular or compressed axonemes (n = 52 for wild type, n = 48 for FAP206-KO). Scale bars, 20 μm (A, B, and E), 1 mm (D), 0.2 μm (F).

Mentions: To localize FAP206 under native conditions of expression, we tagged the FAP206 gene with a sequence encoding a C-terminal green fluorescent protein (GFP). The gross phenotype of the FAP206-GFP strain appeared normal. In both live (Figure 1A and Supplemental Movie S1) and detergent-treated (Triton X-100) cells (Figure 1B), FAP206-GFP was detected exclusively in cilia, where it was distributed uniformly. The detergent resistance indicates that FAP206 is stably associated with the axoneme, in agreement with published biochemical studies (Pazour et al., 2005; Lin et al., 2011; Gupta et al., 2012).


FAP206 is a microtubule-docking adapter for ciliary radial spoke 2 and dynein c.

Vasudevan KK, Song K, Alford LM, Sale WS, Dymek EE, Smith EF, Hennessey T, Joachimiak E, Urbanska P, Wloga D, Dentler W, Nicastro D, Gaertig J - Mol. Biol. Cell (2014)

FAP206 localizes to the ciliary axoneme, and knockout of FAP206 results in cilia-related defects. (A) TIRF image of a live cell expressing FAP206-GFP under the native promoter. (B) A wild-type (negative control) cell (left) and a cell expressing FAP206-GFP under the native promoter (right) extracted with Triton X-100, fixed with paraformaldehyde, and imaged for GFP using a confocal microscope. (C) Culture growth rates for a wild-type CU428 and an FAP206-KO strain. Each data point represents an average for three independent experiments. (D) Paths of swimming wild-type and FAP206-KO cells recorded for 1 s. The average swim velocities were 170 μm/s for the wild type and 50 μm/s for FAP206-KO. (E) Immunofluorescence images of tubulin for wild-type and FAP206-KO cells. For each genotype, an interphase (left) and a dividing cell (right) are shown. (F) Classical TEM images of cilia cross-sections that are either circular (left) or compressed (right). The percentages represent fractions of either circular or compressed axonemes (n = 52 for wild type, n = 48 for FAP206-KO). Scale bars, 20 μm (A, B, and E), 1 mm (D), 0.2 μm (F).
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Related In: Results  -  Collection

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Figure 1: FAP206 localizes to the ciliary axoneme, and knockout of FAP206 results in cilia-related defects. (A) TIRF image of a live cell expressing FAP206-GFP under the native promoter. (B) A wild-type (negative control) cell (left) and a cell expressing FAP206-GFP under the native promoter (right) extracted with Triton X-100, fixed with paraformaldehyde, and imaged for GFP using a confocal microscope. (C) Culture growth rates for a wild-type CU428 and an FAP206-KO strain. Each data point represents an average for three independent experiments. (D) Paths of swimming wild-type and FAP206-KO cells recorded for 1 s. The average swim velocities were 170 μm/s for the wild type and 50 μm/s for FAP206-KO. (E) Immunofluorescence images of tubulin for wild-type and FAP206-KO cells. For each genotype, an interphase (left) and a dividing cell (right) are shown. (F) Classical TEM images of cilia cross-sections that are either circular (left) or compressed (right). The percentages represent fractions of either circular or compressed axonemes (n = 52 for wild type, n = 48 for FAP206-KO). Scale bars, 20 μm (A, B, and E), 1 mm (D), 0.2 μm (F).
Mentions: To localize FAP206 under native conditions of expression, we tagged the FAP206 gene with a sequence encoding a C-terminal green fluorescent protein (GFP). The gross phenotype of the FAP206-GFP strain appeared normal. In both live (Figure 1A and Supplemental Movie S1) and detergent-treated (Triton X-100) cells (Figure 1B), FAP206-GFP was detected exclusively in cilia, where it was distributed uniformly. The detergent resistance indicates that FAP206 is stably associated with the axoneme, in agreement with published biochemical studies (Pazour et al., 2005; Lin et al., 2011; Gupta et al., 2012).

Bottom Line: A knockout of the conserved ciliary protein FAP206 in the ciliate Tetrahymena resulted in slow cell motility.Occasionally, RS2 assembled but lacked both the front prong of its microtubule base and dynein c, whose tail is attached to the front prong.Overexpressed GFP-FAP206 decorated nonciliary microtubules in vivo.

View Article: PubMed Central - PubMed

Affiliation: Department of Cellular Biology, University of Georgia, Athens, GA 30602.

Show MeSH
Related in: MedlinePlus