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Vol. 13, Issue 9, 3303-3313, September 2002
Dartmouth College, Department of Biological Sciences, Hanover, New
Hampshire 03755
Ciliary and flagellar motility is regulated by changes in
intraflagellar calcium. However, the molecular mechanism by which calcium controls motility is unknown. We tested the hypothesis that
calcium regulates motility by controlling dynein-driven microtubule sliding and that the central pair and radial spokes are involved in
this regulation. We isolated axonemes from Chlamydomonas
mutants and measured microtubule sliding velocity in buffers containing 1 mM ATP and various concentrations of calcium. In buffers with pCa > 8, microtubule sliding velocity in axonemes lacking the central apparatus (pf18 and pf15) was
reduced compared with that of wild-type axonemes. In contrast,
at pCa4, dynein activity in pf18 and pf15
axonemes was restored to wild-type level. The calcium-induced increase
in dynein activity in pf18 axonemes was inhibited by antagonists of calmodulin and calmodulin-dependent kinase II. Axonemes
lacking the C1 central tubule (pf16) or lacking radial spoke components (pf14 and pf17) do not
exhibit calcium-induced increase in dynein activity in pCa4 buffer. We
conclude that calcium regulation of flagellar motility involves
regulation of dynein-driven microtubule sliding, that calmodulin and
calmodulin-dependent kinase II may mediate the calcium signal, and that
the central apparatus and radial spokes are key components of the
calcium signaling pathway.
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