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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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Deletion of FAP206 leads to loss of RS2 and associated dynein c in the 96-nm repeat. Isosurface renderings (A–F, K, L) and tomographic slices (G–J) show the averaged 96-nm axonemal repeats of wild type (A, C, D, G, H, K) and FAP206-KO (B, E, F, I, J, L) in longitudinal (A, B, D, F, H, J–L), cross-sectional (C, E, G, I), and bottom views looking from the central pair toward the doublet microtubule (D, F, H, J–L); the dotted line in G indicates the orientation of the tomographic slice shown in H and J. There are three radial spokes: RS1 (green), RS2 (blue), and RS3 (orange) in wild type (A), whereas RS2 is missing in FAP206-KO (B, E, I). The RS2 base is composed of three regions: front (red), back (yellow), and side prongs (light blue and/or black arrowheads). These three regions are connected with each other and form the attachment of RS2 to the doublet A-tubule (At). The back prong of RS2 and the RS3 base (yellow) were previously identified as parts of the CSC (Heuser et al., 2012b). IDA c (pink), which anchors with its tail (pink arrowhead in H) to the A-tubule through the front prong of the RS2 base in WT, is also missing in FAP206-KO (white labels in J, where the IDA c with tail should be). (K, L) Densities representing radial spoke heads and stems were removed to visualize the microtubule-attachment sites of the spoke bases. Note that in the FAP206-KO mutant, the main difference from wild type is the complete absence of the RS2 front prong (red), RS2 back prong (yellow), and dynein c; in contrast, RS1, RS3, and the RS2 side prong (light blue, black arrowhead) appear unaffected in FAP206-KO. All structures shown are subtomogram averages without prior classification analysis. Scale bar, 10 nm.
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Figure 2: Deletion of FAP206 leads to loss of RS2 and associated dynein c in the 96-nm repeat. Isosurface renderings (A–F, K, L) and tomographic slices (G–J) show the averaged 96-nm axonemal repeats of wild type (A, C, D, G, H, K) and FAP206-KO (B, E, F, I, J, L) in longitudinal (A, B, D, F, H, J–L), cross-sectional (C, E, G, I), and bottom views looking from the central pair toward the doublet microtubule (D, F, H, J–L); the dotted line in G indicates the orientation of the tomographic slice shown in H and J. There are three radial spokes: RS1 (green), RS2 (blue), and RS3 (orange) in wild type (A), whereas RS2 is missing in FAP206-KO (B, E, I). The RS2 base is composed of three regions: front (red), back (yellow), and side prongs (light blue and/or black arrowheads). These three regions are connected with each other and form the attachment of RS2 to the doublet A-tubule (At). The back prong of RS2 and the RS3 base (yellow) were previously identified as parts of the CSC (Heuser et al., 2012b). IDA c (pink), which anchors with its tail (pink arrowhead in H) to the A-tubule through the front prong of the RS2 base in WT, is also missing in FAP206-KO (white labels in J, where the IDA c with tail should be). (K, L) Densities representing radial spoke heads and stems were removed to visualize the microtubule-attachment sites of the spoke bases. Note that in the FAP206-KO mutant, the main difference from wild type is the complete absence of the RS2 front prong (red), RS2 back prong (yellow), and dynein c; in contrast, RS1, RS3, and the RS2 side prong (light blue, black arrowhead) appear unaffected in FAP206-KO. All structures shown are subtomogram averages without prior classification analysis. Scale bar, 10 nm.

Mentions: Previous biochemical and genetic studies linked FAP206 to the 96-nm outer doublet repeat (Lin et al., 2011; Gupta et al., 2012). We used cryo–electron tomography and subtomogram averaging to compare the high-resolution three-dimensional (3D) structure of the 96-nm repeat between the wild-type and FAP206-KO axonemes. We reconstructed cryotomograms of 10 wild-type and 7 FAP206-KO axonemes and averaged 1300 and 800 axonemal repeats, respectively. The most striking difference in the subtomogram averages of all repeats is the absence of the middle radial spoke RS2 and its closely associated IDA, dynein c, in the FAP206-KO axonemes (Figure 2 and Supplemental Movies S4 and S5). In the wild-type 96-nm repeat, the RS2 base is attached to the A-tubule by three contacts: the front, back, and side prongs (Figure 2, C, D, H, and K). The front and back prongs (red and yellow in Figure 2, respectively) resemble the corresponding structures in Chlamydomonas (Barber et al., 2012; Lin et al., 2012), whereas the side prong (light blue in Figure 2) is, so far, unique to RS2 of Tetrahymena. In the axonemal repeat of FAP206-KO, the front and back prong, of RS2 are absent, whereas the side prong remains (Figure 2, E, F, J, and L, and Supplemental Movie S6). We conclude that the base of the Tetrahymena RS2 attaches to the microtubule with three prongs, and the assembly of two of these prongs (front and back) depends on FAP206.


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)

Deletion of FAP206 leads to loss of RS2 and associated dynein c in the 96-nm repeat. Isosurface renderings (A–F, K, L) and tomographic slices (G–J) show the averaged 96-nm axonemal repeats of wild type (A, C, D, G, H, K) and FAP206-KO (B, E, F, I, J, L) in longitudinal (A, B, D, F, H, J–L), cross-sectional (C, E, G, I), and bottom views looking from the central pair toward the doublet microtubule (D, F, H, J–L); the dotted line in G indicates the orientation of the tomographic slice shown in H and J. There are three radial spokes: RS1 (green), RS2 (blue), and RS3 (orange) in wild type (A), whereas RS2 is missing in FAP206-KO (B, E, I). The RS2 base is composed of three regions: front (red), back (yellow), and side prongs (light blue and/or black arrowheads). These three regions are connected with each other and form the attachment of RS2 to the doublet A-tubule (At). The back prong of RS2 and the RS3 base (yellow) were previously identified as parts of the CSC (Heuser et al., 2012b). IDA c (pink), which anchors with its tail (pink arrowhead in H) to the A-tubule through the front prong of the RS2 base in WT, is also missing in FAP206-KO (white labels in J, where the IDA c with tail should be). (K, L) Densities representing radial spoke heads and stems were removed to visualize the microtubule-attachment sites of the spoke bases. Note that in the FAP206-KO mutant, the main difference from wild type is the complete absence of the RS2 front prong (red), RS2 back prong (yellow), and dynein c; in contrast, RS1, RS3, and the RS2 side prong (light blue, black arrowhead) appear unaffected in FAP206-KO. All structures shown are subtomogram averages without prior classification analysis. Scale bar, 10 nm.
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Related In: Results  -  Collection

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Figure 2: Deletion of FAP206 leads to loss of RS2 and associated dynein c in the 96-nm repeat. Isosurface renderings (A–F, K, L) and tomographic slices (G–J) show the averaged 96-nm axonemal repeats of wild type (A, C, D, G, H, K) and FAP206-KO (B, E, F, I, J, L) in longitudinal (A, B, D, F, H, J–L), cross-sectional (C, E, G, I), and bottom views looking from the central pair toward the doublet microtubule (D, F, H, J–L); the dotted line in G indicates the orientation of the tomographic slice shown in H and J. There are three radial spokes: RS1 (green), RS2 (blue), and RS3 (orange) in wild type (A), whereas RS2 is missing in FAP206-KO (B, E, I). The RS2 base is composed of three regions: front (red), back (yellow), and side prongs (light blue and/or black arrowheads). These three regions are connected with each other and form the attachment of RS2 to the doublet A-tubule (At). The back prong of RS2 and the RS3 base (yellow) were previously identified as parts of the CSC (Heuser et al., 2012b). IDA c (pink), which anchors with its tail (pink arrowhead in H) to the A-tubule through the front prong of the RS2 base in WT, is also missing in FAP206-KO (white labels in J, where the IDA c with tail should be). (K, L) Densities representing radial spoke heads and stems were removed to visualize the microtubule-attachment sites of the spoke bases. Note that in the FAP206-KO mutant, the main difference from wild type is the complete absence of the RS2 front prong (red), RS2 back prong (yellow), and dynein c; in contrast, RS1, RS3, and the RS2 side prong (light blue, black arrowhead) appear unaffected in FAP206-KO. All structures shown are subtomogram averages without prior classification analysis. Scale bar, 10 nm.
Mentions: Previous biochemical and genetic studies linked FAP206 to the 96-nm outer doublet repeat (Lin et al., 2011; Gupta et al., 2012). We used cryo–electron tomography and subtomogram averaging to compare the high-resolution three-dimensional (3D) structure of the 96-nm repeat between the wild-type and FAP206-KO axonemes. We reconstructed cryotomograms of 10 wild-type and 7 FAP206-KO axonemes and averaged 1300 and 800 axonemal repeats, respectively. The most striking difference in the subtomogram averages of all repeats is the absence of the middle radial spoke RS2 and its closely associated IDA, dynein c, in the FAP206-KO axonemes (Figure 2 and Supplemental Movies S4 and S5). In the wild-type 96-nm repeat, the RS2 base is attached to the A-tubule by three contacts: the front, back, and side prongs (Figure 2, C, D, H, and K). The front and back prongs (red and yellow in Figure 2, respectively) resemble the corresponding structures in Chlamydomonas (Barber et al., 2012; Lin et al., 2012), whereas the side prong (light blue in Figure 2) is, so far, unique to RS2 of Tetrahymena. In the axonemal repeat of FAP206-KO, the front and back prong, of RS2 are absent, whereas the side prong remains (Figure 2, E, F, J, and L, and Supplemental Movie S6). We conclude that the base of the Tetrahymena RS2 attaches to the microtubule with three prongs, and the assembly of two of these prongs (front and back) depends on FAP206.

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