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Vol. 14, Issue 2, 774-785, February 2003

Identification of a Novel Leucine-rich Repeat Protein as a Component of Flagellar Radial Spoke in the Ascidian Ciona intestinalis

Potturi Padma,^* Yuhkoh Satouh,^* Ken-ichi Wakabayashi,^§ Akiko Hozumi,^* Yuji Ushimaru,^* Ritsu Kamiya, and Kazuo Inaba^*¶

 ^*Asamushi Marine Biological Station, Graduate School of Science, Tohoku University, Aomori 039-3501, Japan;  National Institute for Basic Biology, Okazaki 444-8585, Japan; and  Department of Biological Sciences, Graduate School of Science, University of Tokyo, Tokyo 113-0033, Japan

Axonemes are highly organized microtubule-based structures conserved in many eukaryotes. In an attempt to study axonemes by a proteomics approach, we selectively cloned cDNAs of axonemal proteins by immunoscreening the testis cDNA library from the ascidian Ciona intestinalis by using an antiserum against whole axonemes. We report here a 37-kDa protein of which cDNA occurred most frequently among total positive clones. This protein, named LRR37, belongs to the class of SDS22+ leucine-rich repeat (LRR) family. LRR37 is different from the LRR outer arm dynein light chain reported in Chlamydomonas and sea urchin flagella, and thus represents a novel axonemal LRR protein. Immunoelectron microscopy by using a polyclonal antibody against LRR37 showed that it is localized on the tip of the radial spoke, most likely on the spoke head. The LRR37 protein in fact seems to form a complex together with radial spoke protein 3 in a KI extract of the axonemes. These results suggest that LRR37 is a component of the radial spoke head and is involved in the interaction with other radial spoke components or proteins in the central pair projection.

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P.P. and Y.S. contributed equally to this work.

^§ Present address: Department of Biochemistry, University of Connecticut Health Center, Farmington, CT 06030-3305.

^¶ Corresponding author. E-mail address: inaba{at}biology.tohoku.ac.jp.

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Molecular Biology of the Cell
Vol. 14, 774-785, February 2003
Copyright © 2003 by The American Society for Cell Biology

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