Outer Dense Fiber 2 Is a Widespread Centrosome Scaffold Component Preferentially Associated with Mother Centrioles: Its Identification from Isolated Centrosomes

Abstract

Because centrosomes were enriched in the bile canaliculi fraction from the chicken liver through their association with apical membranes, we developed a procedure for isolation of centrosomes from this fraction. With the use of the centrosomes, we generated centrosome-specific monoclonal antibodies. Three of the monoclonal antibodies recognized an antigen of ~90 kDa. Cloning of its cDNA identified this antigen as a chicken homologue of outer dense fiber 2 protein (Odf2), which was initially identified as a sperm outer dense fiber-specific component. Exogenously expressed and endogenous Odf2 were shown to be concentrated at the centrosomes in a microtubule-independent manner in various types of cells at both light and electron microscopic levels. Odf2 exhibited a cell cycle-dependent pattern of localization and was preferentially associated with the mother centrioles in G0/G1-phase. Toward G1/S-phase before centrosome duplication, it became detectable in both mother and daughter centrioles. In the isolated bile canaliculi and centrosomes, Odf2, in contrast to other centrosomal components, was highly resistant to KI extraction. These findings indicate that Odf2 is a widespread KI-insoluble scaffold component of the centrosome matrix, which may be involved in the maturation event of daughter centrioles.

A gene trap knockout of the abundant sperm tail protein, outer dense fiber 2, results in preimplantation lethality

“…However we also found low Odf2 expression in all adult tissues examined. This finding is consistent with more recent studies that have identified ODF2 as a widespread component of the centrosome (Nakagawa et al, 2001). Furthermore, F9 mouse cells with a homozygous targeted knockout of Odf2 displayed centrosomes that failed to generate primary cilia, implying that Odf2 gene function is not restricted to formation of the outer dense fibers of the sperm tail, but rather plays an additional wide-spread role at the centrosome (Ishikawa et al, 2005).…”

“…However, in a recent publication, monoclonal antibodies directed against purified chicken centrosomes have identified ODF2 protein as a widespread centrosomal component (Nakagawa et al, 2001). Further studies have also suggested that ODF2 protein at the centrosome is able to associate both with itself and with microtubules (Donkor et al, 2004).…”

The deubiquitinating enzyme CYLD controls apical docking of basal bodies in ciliated epithelial cells

“…Furthermore, centriolar satellites are believed to drive cargo to the centrosome and basal bodies 35 , and are also required for ciliogenesis 36 . To ascertain the presence of centriolar satellites and appendages, we stained ependymal cells with antibodies directed against PCM1 and OFD1, two proteins of centriolar satellites 35,36 , as well as with antibodies against Cep164 and CCDC123 (Cep123), two proteins located within the distal appendages 37,38 , and ODF2, a protein located both at distal and subdistal appendages 39 . All antibodies decorated centriolar satellites or basal bodies irrespective of the genotype (Fig.…”

Centrosomal nucleolin is required for microtubule network organization

“…Cenexin is localized on these 2 types of appendages 35 and is involved in both ciliogenesis and the stabilization of centrosomal microtubules. 13 It remains to be determined if nucleolin could also have a function related to ciliogenesis.…”

“…1), we then determined whether it was randomly distributed on the 2 centrioles, or if it was specifically associated with the centriole carrying mature or immature markers. Nucleolin localization was assessed by performing co-immunodetections with distal (CEP164), 33 subdistal (ninein) 34 or both distal and subdistal (cenexin) 35 appendage proteins (Fig. 3).…”