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Two appendages homologous between basal bodies and centrioles are formed using distinct Odf2 domains.

Tateishi K, Yamazaki Y, Nishida T, Watanabe S, Kunimoto K, Ishikawa H, Tsukita S - J. Cell Biol. (2013)

Bottom Line: We examined the effects of Odf2 deletion constructs on ciliogenesis in Odf2-knockout F9 cells.We used the differential formation of appendages by Odf2 deletion constructs to study the incorporation and function of molecules associated with each appendage type.We found that transition fibers and distal appendages were required for ciliogenesis and subdistal appendages stabilized the centrosomal microtubules.

View Article: PubMed Central - HTML - PubMed

Affiliation: Laboratory of Biological Science, Graduate School of Frontier Biosciences and Graduate School of Medicine, Osaka University, Osaka 565-0871, Japan.

ABSTRACT
Ciliogenesis is regulated by context-dependent cellular cues, including some transduced through appendage-like structures on ciliary basal bodies called transition fibers and basal feet. However, the molecular basis for this regulation is not fully understood. The Odf2 gene product, ODF2/cenexin, is essential for both ciliogenesis and the formation of the distal and subdistal appendages on centrioles, which become basal bodies. We examined the effects of Odf2 deletion constructs on ciliogenesis in Odf2-knockout F9 cells. Electron microscopy revealed that ciliogenesis and transition fiber formation required the ODF2/cenexin fragment containing amino acids (aa) 188-806, whereas basal foot formation required aa 1-59 and 188-806. These sequences also formed distal and subdistal appendages, respectively, indicating that the centriole appendages are molecularly analogous to those on basal bodies. We used the differential formation of appendages by Odf2 deletion constructs to study the incorporation and function of molecules associated with each appendage type. We found that transition fibers and distal appendages were required for ciliogenesis and subdistal appendages stabilized the centrosomal microtubules.

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Reconstitution of centriole appendages by Odf2 deletion constructs. (A) Electron micrographs showing distal appendages (DAs) and/or subdistal appendages (SAs) on centrioles. Thin-section electron microscopic images show cross sections (Cross) and longitudinal sections (Longitudinal) of centrioles. More than five samples were analyzed in each case. Tomography, UHVEMT images of centrioles; DAs, blue arrows; SAs, red arrowheads; WT, (DA+SA+) centriole in WT F9 cells; Δ4/5, (DA+SA−) centriole in Δ4/5 construct–expressing Odf2-KO F9 cells; Δ6/7, (DA+SA+) centriole in Δ6/7 construct–expressing Odf2-KO F9 cells; KO, (DA−SA−) centriole; TFs, blue arrows; BF, red arrowheads. Bars, 0.2 µm. Insets, schematic drawings of electron microscopic images of centrioles. (B) Summary of findings on the reconstitution of the basal body/centriole appendages.
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fig2: Reconstitution of centriole appendages by Odf2 deletion constructs. (A) Electron micrographs showing distal appendages (DAs) and/or subdistal appendages (SAs) on centrioles. Thin-section electron microscopic images show cross sections (Cross) and longitudinal sections (Longitudinal) of centrioles. More than five samples were analyzed in each case. Tomography, UHVEMT images of centrioles; DAs, blue arrows; SAs, red arrowheads; WT, (DA+SA+) centriole in WT F9 cells; Δ4/5, (DA+SA−) centriole in Δ4/5 construct–expressing Odf2-KO F9 cells; Δ6/7, (DA+SA+) centriole in Δ6/7 construct–expressing Odf2-KO F9 cells; KO, (DA−SA−) centriole; TFs, blue arrows; BF, red arrowheads. Bars, 0.2 µm. Insets, schematic drawings of electron microscopic images of centrioles. (B) Summary of findings on the reconstitution of the basal body/centriole appendages.

Mentions: We first examined the centrosomes in cells expressing the Odf2 ciliogenic constructs that formed TFs, and found that these constructs also formed the centriole DAs. In contrast, the nonciliogenic constructs did not reconstitute the DAs (Fig. 2, Fig. 3, Fig. S1, Fig. S3 B, and Videos 5–9). Thus, the same domains of Odf2 were required to form ciliary TFs and centriole DAs, suggesting that these structures had some molecular architecture and function in common.


Two appendages homologous between basal bodies and centrioles are formed using distinct Odf2 domains.

Tateishi K, Yamazaki Y, Nishida T, Watanabe S, Kunimoto K, Ishikawa H, Tsukita S - J. Cell Biol. (2013)

Reconstitution of centriole appendages by Odf2 deletion constructs. (A) Electron micrographs showing distal appendages (DAs) and/or subdistal appendages (SAs) on centrioles. Thin-section electron microscopic images show cross sections (Cross) and longitudinal sections (Longitudinal) of centrioles. More than five samples were analyzed in each case. Tomography, UHVEMT images of centrioles; DAs, blue arrows; SAs, red arrowheads; WT, (DA+SA+) centriole in WT F9 cells; Δ4/5, (DA+SA−) centriole in Δ4/5 construct–expressing Odf2-KO F9 cells; Δ6/7, (DA+SA+) centriole in Δ6/7 construct–expressing Odf2-KO F9 cells; KO, (DA−SA−) centriole; TFs, blue arrows; BF, red arrowheads. Bars, 0.2 µm. Insets, schematic drawings of electron microscopic images of centrioles. (B) Summary of findings on the reconstitution of the basal body/centriole appendages.
© Copyright Policy - openaccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC3824012&req=5

fig2: Reconstitution of centriole appendages by Odf2 deletion constructs. (A) Electron micrographs showing distal appendages (DAs) and/or subdistal appendages (SAs) on centrioles. Thin-section electron microscopic images show cross sections (Cross) and longitudinal sections (Longitudinal) of centrioles. More than five samples were analyzed in each case. Tomography, UHVEMT images of centrioles; DAs, blue arrows; SAs, red arrowheads; WT, (DA+SA+) centriole in WT F9 cells; Δ4/5, (DA+SA−) centriole in Δ4/5 construct–expressing Odf2-KO F9 cells; Δ6/7, (DA+SA+) centriole in Δ6/7 construct–expressing Odf2-KO F9 cells; KO, (DA−SA−) centriole; TFs, blue arrows; BF, red arrowheads. Bars, 0.2 µm. Insets, schematic drawings of electron microscopic images of centrioles. (B) Summary of findings on the reconstitution of the basal body/centriole appendages.
Mentions: We first examined the centrosomes in cells expressing the Odf2 ciliogenic constructs that formed TFs, and found that these constructs also formed the centriole DAs. In contrast, the nonciliogenic constructs did not reconstitute the DAs (Fig. 2, Fig. 3, Fig. S1, Fig. S3 B, and Videos 5–9). Thus, the same domains of Odf2 were required to form ciliary TFs and centriole DAs, suggesting that these structures had some molecular architecture and function in common.

Bottom Line: We examined the effects of Odf2 deletion constructs on ciliogenesis in Odf2-knockout F9 cells.We used the differential formation of appendages by Odf2 deletion constructs to study the incorporation and function of molecules associated with each appendage type.We found that transition fibers and distal appendages were required for ciliogenesis and subdistal appendages stabilized the centrosomal microtubules.

View Article: PubMed Central - HTML - PubMed

Affiliation: Laboratory of Biological Science, Graduate School of Frontier Biosciences and Graduate School of Medicine, Osaka University, Osaka 565-0871, Japan.

ABSTRACT
Ciliogenesis is regulated by context-dependent cellular cues, including some transduced through appendage-like structures on ciliary basal bodies called transition fibers and basal feet. However, the molecular basis for this regulation is not fully understood. The Odf2 gene product, ODF2/cenexin, is essential for both ciliogenesis and the formation of the distal and subdistal appendages on centrioles, which become basal bodies. We examined the effects of Odf2 deletion constructs on ciliogenesis in Odf2-knockout F9 cells. Electron microscopy revealed that ciliogenesis and transition fiber formation required the ODF2/cenexin fragment containing amino acids (aa) 188-806, whereas basal foot formation required aa 1-59 and 188-806. These sequences also formed distal and subdistal appendages, respectively, indicating that the centriole appendages are molecularly analogous to those on basal bodies. We used the differential formation of appendages by Odf2 deletion constructs to study the incorporation and function of molecules associated with each appendage type. We found that transition fibers and distal appendages were required for ciliogenesis and subdistal appendages stabilized the centrosomal microtubules.

Show MeSH
Related in: MedlinePlus