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J. Biol. Chem., Vol. 280, Issue 50, 41412-41420, December 16, 2005

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An Axonemal Dynein Particularly Important for Flagellar Movement at High Viscosity

IMPLICATIONS FROM A NEW CHLAMYDOMONAS MUTANT DEFICIENT IN THE DYNEIN HEAVY CHAIN GENE DHC9^*

Toshiki Yagi, Itsushi Minoura1, Akiko Fujiwara, Ryo Saito¶, Takuo Yasunaga¶, Masafumi Hirono, and Ritsu Kamiya||2

From the Department of Biological Sciences, University of Tokyo, 7-3-1 Hongo, Tokyo 113-0033, the National Institute for Basic Biology, Okazaki 444-8585, the ^¶Department of Biochemical Engineering and Science, Kyushu Institute of Technology, Iizuka 820-8502, and ^||CREST, Japan Science and Technology Corporation, Kawaguchi 332-0012, Japan

Ciliary and flagellar axonemes contain multiple inner arm dyneins of which the functional difference is largely unknown. In this study, a Chlamydomonas mutant, ida9, lacking inner arm dynein c was isolated and shown to carry a mutation in the DHC9 dynein heavy chain gene. The cDNA sequence of DHC9 was determined, and its information was used to show that >80% of it is lost in the mutant. Electron microscopy and image analysis showed that the ida9 axoneme lacked electron density near the base of the S2 radial spoke, indicating that dynein c localizes to this site. The mutant ida9 swam only slightly slower than the wild type in normal media. However, swimming velocity was greatly reduced when medium viscosity was modestly increased. Thus, dynein c in wild type axonemes must produce a significant force when flagella are beating in viscous media. Because motility analyses in vitro have shown that dynein c is the fastest among all the inner arm dyneins, we can regard this dynein as a fast yet powerful motor.

Received for publication, August 17, 2005 , and in revised form, October 12, 2005.

^* This work was supported by a grant from the Ministry of Education, Culture, Sports, Science, and Technology. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The on-line version of this article (available at http://www.jbc.org) contains supplemental Figs. S1 and S2.

¹ Present address: RIKEN Brain Science Institute, Saitama, Wako-shi 351-0198, Japan.

² To whom correspondence should be addressed: Dept. of Biological Sciences, Graduate School of Science, University of Tokyo, Tokyo 113-0033, Japan. Tel./Fax: 81-3-5841-4632; E-mail: kamiyar{at}biol.s.u-tokyo.ac.jp.

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