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Vol. 12, Issue 6, 1687-1697, June 2001
and§
*Department of Cell Biology, Faculty of Medicine, Kyoto University,
Sakyo-ku, Kyoto 606-8501, Japan; 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.
Third Department of
Internal Medicine, Kyoto Prefectural University of Medicine,
Kamigyo-ku, Kyoto 602-8566, Japan; §College of Medical
Technology, Kyoto University, Sakyo-ku, Kyoto 606-8501, Japan; and
Tsukita Cell Axis Project, Exploratory Research for
Advanced Technology, Japan Science and Technology Corporation, Kyoto
Research Park, Shimogyo-ku, Kyoto 600-8813, Japan
Corresponding author. E-mail address:
atsukita{at}mfour.med.kyoto-u.ac.jp.
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