Limits...
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.

Show MeSH

Related in: MedlinePlus

Immunofluorescence microscopic images of centriole appendages in the DA+SA+, DA-SA−, and DA+SA− patterns. (A) Immunofluorescence for ninein, centriolin, CEP164, and OFD1. WT, (DA+SA+) centriole in WT F9 cells; KO, (DA-SA−) centriole in Odf2-KO 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. (B) Schematic drawing showing the localization of centrosomal components. MC, mother centriole; DC, daughter centriole. Bars, 500 nm.
© Copyright Policy - openaccess
Related In: Results  -  Collection

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

fig4: Immunofluorescence microscopic images of centriole appendages in the DA+SA+, DA-SA−, and DA+SA− patterns. (A) Immunofluorescence for ninein, centriolin, CEP164, and OFD1. WT, (DA+SA+) centriole in WT F9 cells; KO, (DA-SA−) centriole in Odf2-KO 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. (B) Schematic drawing showing the localization of centrosomal components. MC, mother centriole; DC, daughter centriole. Bars, 500 nm.

Mentions: The two sets of homologous appendages on basal bodies and centrioles appear to be molecularly distinct. Their dependence on different domains of the Odf2 gene product could result in the distinct sets of associated molecules incorporated into the different appendage types. The three patterns of reconstitution (i.e., centrioles bearing no appendages, only DAs, or DAs and SAs, depending on the sequences contained in the Odf2 deletion construct) provided a powerful tool for confirming which molecular components are associated with DAs or SAs, respectively. We therefore examined the immunofluorescence signals for DA- and SA-associated proteins (ninein, centriolin, and CEP164) and centriole proteins (chibby and OFD1) in their respective reconstituted appendages. To examine the relative spatial relationships of the appendages in centrosomes, we stained the centrosomes with anti-GFP to visualize GFP-tagged ODF2/cenexin and with anti–γ-tubulin to label centrosomes (Fig. 4).


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)

Immunofluorescence microscopic images of centriole appendages in the DA+SA+, DA-SA−, and DA+SA− patterns. (A) Immunofluorescence for ninein, centriolin, CEP164, and OFD1. WT, (DA+SA+) centriole in WT F9 cells; KO, (DA-SA−) centriole in Odf2-KO 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. (B) Schematic drawing showing the localization of centrosomal components. MC, mother centriole; DC, daughter centriole. Bars, 500 nm.
© Copyright Policy - openaccess
Related In: Results  -  Collection

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

fig4: Immunofluorescence microscopic images of centriole appendages in the DA+SA+, DA-SA−, and DA+SA− patterns. (A) Immunofluorescence for ninein, centriolin, CEP164, and OFD1. WT, (DA+SA+) centriole in WT F9 cells; KO, (DA-SA−) centriole in Odf2-KO 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. (B) Schematic drawing showing the localization of centrosomal components. MC, mother centriole; DC, daughter centriole. Bars, 500 nm.
Mentions: The two sets of homologous appendages on basal bodies and centrioles appear to be molecularly distinct. Their dependence on different domains of the Odf2 gene product could result in the distinct sets of associated molecules incorporated into the different appendage types. The three patterns of reconstitution (i.e., centrioles bearing no appendages, only DAs, or DAs and SAs, depending on the sequences contained in the Odf2 deletion construct) provided a powerful tool for confirming which molecular components are associated with DAs or SAs, respectively. We therefore examined the immunofluorescence signals for DA- and SA-associated proteins (ninein, centriolin, and CEP164) and centriole proteins (chibby and OFD1) in their respective reconstituted appendages. To examine the relative spatial relationships of the appendages in centrosomes, we stained the centrosomes with anti-GFP to visualize GFP-tagged ODF2/cenexin and with anti–γ-tubulin to label centrosomes (Fig. 4).

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