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Journal of Biochemistry Advance Access originally published online on August 26, 2006
Journal of Biochemistry 2006 140(4):525-533; doi:10.1093/jb/mvj182
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© 2006 The Japanese Biochemical Society.

ARTICLE

Homologues of Radial Spoke Head Proteins Interact with Ca2+/Calmodulin in Tetrahymena Cilia

Hironori Ueno*, Yoshinori Iwataki and Osamu Numata{dagger}

Graduate School of Life and Environmental Sciences, University of Tsukuba, Tsukuba, Ibaraki 305-8572

{dagger} To whom correspondence should be addressed. Tel: +81-29-853-6648, Fax: +81-29-853-6614, E-mail numata{at}sakura.cc.tsukuba.ac.jp

Calmodulin (CaM) is an axonemal component. To examine the pathway of Ca2+/CaM signaling in cilia, using Ca2+/CaM-affinity column, we identified seven Ca2+/CaM-associated proteins from a crude dynein fraction and isolated 62 kDa (p62) and 66 kDa (p66) Ca2+/CaM-associated proteins in Tetrahymena cilia. The amino acid sequences deduced from the p62 and p66 cDNA sequences suggested that these proteins were similar to Chlamydomonas radial spoke proteins 4 and 6 (RSP4 and RSP6), components of the radial spoke head, and sea urchin sperm p63, which is a homologue of RSP4/6, and isolated as a key component that affect flagellar bending patterns. Although p62 and p66 do not have a conventional CaM-binding site, those have consecutive sequences which showed high normalized scores (≥ 5) from a CaM target database. These consecutive sequences were also found in RSP4, RSP6, and p63. These radial spoke heads proteins have a high similarity region composed of 15 amino acids between the five proteins. Immunoelectron microscopy using anti-CaM antibody showed that CaM was localized along the outer edge of the curved central pair microtubules in axoneme. Therefore, it is possible that the interaction between Ca2+/CaM and radial spoke head control axonemal curvature in the ciliary and flagellar waveform.

* Present address: Research Institute for Cell Engineering, National Institute for Advanced Industrial Science and Technology, Tsukuba Central 4, 1-1-1 Higashi, Tsukuba, Ibaraki 305-8562.


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