title: Casein kinase I is anchored on axonemal doublet microtubules and regulates flagellar dynein phosphorylation and activity.
abstract: 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.
authors.affiliation: Department of Cell Biology, Emory University School of Medicine, Atlanta, Georgia 30322, USA.
authors.forename: P
authors.initials: P
authors.lastname: Yang
authors.forename: W S
authors.initials: WS
authors.lastname: Sale
mesh_terms: D047388:Casein Kinases; D003599:Cytoskeleton / Q000201:enzymology; D004398:Dyneins / Q000737:chemistry / Q000378:metabolism*; D005407:Flagella / Q000201:enzymology*; D008870:Microtubules / Q000201:enzymology*; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism*
chemical_list: D011494:Protein Kinases; D047388:Casein Kinases; D004398:Dyneins