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J. Biol. Chem., Vol. 275, Issue 25, 18905-18912, June 23, 2000
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From the Department of Cell Biology, Emory University School of
Medicine, Atlanta, Georgia 30322
Flagellar dynein activity is regulated by
phosphorylation. One critical phosphoprotein substrate in
Chlamydomonas is the 138-kDa intermediate chain (IC138) of
the inner arm dyneins (Habermacher, G., and Sale, W. S. (1997)
J. Cell Biol. 136, 167-176). In this study, several
approaches were used to determine that casein kinase I (CKI) is
physically anchored in the flagellar axoneme and regulates IC138
phosphorylation and dynein activity. First, using a videomicroscopic motility assay, selective CKI inhibitors rescued dynein-driven microtubule sliding in axonemes isolated from paralyzed flagellar mutants lacking radial spokes. Rescue of dynein activity failed in
axonemes isolated from these mutant cells lacking IC138. Second, CKI
was unequivocally identified in salt extracts from isolated axonemes,
whereas casein kinase II was excluded from the flagellar compartment.
Third, Western blots indicate that within flagella, CKI is anchored
exclusively to the axoneme. Analysis of multiple Chlamydomonas motility mutants suggests that the axonemal
CKI is located on the outer doublet microtubules. Finally, CKI
inhibitors that rescued dynein activity blocked phosphorylation of
IC138. We propose that CKI is anchored on the outer doublet
microtubules in position to regulate flagellar dynein.
Casein Kinase I Is Anchored on Axonemal Doublet Microtubules and
Regulates Flagellar Dynein Phosphorylation and Activity*
*
This work was supported by the March of Dimes Birth Defects
Foundation and National Institutes of Health Grants GM51173 and GM17666.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom correspondence should be addressed: Dept. of Cell Biology,
Emory University School of Medicine, 1648 Pierce Dr., Atlanta, GA
30322. Tel.: 404-727-6265; Fax: 404-727-6256; E-mail:
win@cellbio.emory.edu.
Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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