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Published online 1 June 2004. doi:10.1083/jcb.200402022
The Rockefeller University Press, 0021-9525 $8.00
JCB, Volume 165, Number 5, 663-671
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Article

 Bld10p, a novel protein essential for basal body assembly in Chlamydomonas : localization to the cartwheel, the first ninefold symmetrical structure appearing during assembly



Kumi Matsuura1, Paul A. Lefebvre2,3, Ritsu Kamiya1,4, and Masafumi Hirono1

1 Department of Biological Sciences, University of Tokyo, Tokyo 113-0033, Japan
2 Department of Genetics, Cell Biology and Development
3 Department of Plant Biology, University of Minnesota, St. Paul, MN 55108
4 Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Corporation (JST), Nagoya 450-0003, Japan

Address correspondence to Masafumi Hirono, Dept. of Biological Sciences, University of Tokyo, Bunkyo-ku, Tokyo 113-0033, Japan. Tel.: 81-3-5841-4429. Fax: 81-3-5802-2734. email: hirono{at}biol.s.u-tokyo.ac.jp

How centrioles and basal bodies assemble is a long-standing puzzle in cell biology. To address this problem, we analyzed a novel basal body-defective Chlamydomonas reinhardtii mutant isolated from a collection of flagella-less mutants. This mutant, bld10, displayed disorganized mitotic spindles and cytoplasmic microtubules, resulting in abnormal cell division and slow growth. Electron microscopic observation suggested that bld10 cells totally lack basal bodies. The product of the BLD10 gene (Bld10p) was found to be a novel coiled-coil protein of 170 kD. Immunoelectron microscopy localizes Bld10p to the cartwheel, a structure with ninefold rotational symmetry positioned near the proximal end of the basal bodies. Because the cartwheel forms the base from which the triplet microtubules elongate, we suggest that Bld10p plays an essential role in an early stage of basal body assembly. A viable mutant having such a severe basal body defect emphasizes the usefulness of Chlamydomonas in studying the mechanism of basal body/centriole assembly by using a variety of mutants.

Key Words: centriole; centrosome; mitotic spindle apparatus; flagella; coiled-coil

K. Matsuura's present address is Molecular Membrane Biology Laboratory, RIKEN, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan.

Abbreviations used in this paper: BAC, bacterial artificial chromosome; NFAp, nucleoflagellar apparatus.


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