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Published online 11 June 2001. doi:10.1083/jcb.153.6.1315
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© The Rockefeller University Press, 0021-9525/2001//1315 $5.00
The Journal of Cell Biology, Volume 153, Number 6, , 2001 1315-1326

Original Article

Localization of Calmodulin and Dynein Light Chain Lc8 in Flagellar Radial Spokes



Pinfen Yanga, Dennis R. Dienerb, Joel L. Rosenbaumb, and Winfield S. Salea

a Department of Cell Biology, Emory University, School of Medicine, Atlanta, Georgia 30322
b Department of Molecular, Cellular and Developmental Biology, Yale University, New Haven, Connecticut 06520
Department of Cell Biology, Emory University School of Medicine, 1648 Pierce Dr., Atlanta, GA 30222.(404) 727-6256(404) 727-6265

win{at}cellbio.emory.edu

Genetic and in vitro analyses have revealed that radial spokes play a crucial role in regulation of ciliary and flagellar motility, including control of waveform. However, the mechanisms of regulation are not understood. Here, we developed a novel procedure to isolate intact radial spokes as a step toward understanding the mechanism by which these complexes regulate dynein activity. The isolated radial spokes sediment as 20S complexes that are the size and shape of radial spokes. Extracted radial spokes rescue radial spoke structure when reconstituted with isolated axonemes derived from the radial spoke mutant pf14. Isolated radial spokes are composed of the 17 previously defined spoke proteins as well as at least five additional proteins including calmodulin and the ubiquitous dynein light chain LC8. Analyses of flagellar mutants and chemical cross-linking studies demonstrated calmodulin and LC8 form a complex located in the radial spoke stalk. We postulate that calmodulin, located in the radial spoke stalk, plays a role in calcium control of flagellar bending.

Key Words: calcium • cilia • dynein • flagella • calmodulin

© 2001 The Rockefeller University Press

Abbreviations used in this paper: 2D, two-dimensional; KI, potassium iodide; KP, potassium phosphate; pI, isoelectric point; PKA, protein kinase A.


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