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Hongmin Qin, Dennis R. Diener, Stefan Geimer, Douglas G. Cole, and Joel L. Rosenbaum

Department of Molecular, Cellular, and Developmental Biology, Yale University, New Haven, CT 06520

Address correspondence to Joel Rosenbaum, MCDB Dept., Yale University, New Haven, CT 06520. Tel.: (203) 432-3472. Fax: (203) 432-5059. email: joel.rosenbaum{at}yale.edu

Intraflagellar transport (IFT) is the bidirectional movement of multisubunit protein particles along axonemal microtubules and is required for assembly and maintenance of eukaryotic flagella and cilia. One posited role of IFT is to transport flagellar precursors to the flagellar tip for assembly. Here, we examine radial spokes, axonemal subunits consisting of 22 polypeptides, as potential cargo for IFT. Radial spokes were found to be partially assembled in the cell body, before being transported to the flagellar tip by anterograde IFT. Fully assembled radial spokes, detached from axonemal microtubules during flagellar breakdown or turnover, are removed from flagella by retrograde IFT. Interactions between IFT particles, motors, radial spokes, and other axonemal proteins were verified by coimmunoprecipitation of these proteins from the soluble fraction of Chlamydomonas flagella. These studies indicate that one of the main roles of IFT in flagellar assembly and maintenance is to transport axonemal proteins in and out of the flagellum.

Key Words: cilia; intraflagellar; transport; kinesin; radial spoke

D.G. Cole's present address is Dept. of Microbiology, Molecular Biology and Biochemistry, University of Idaho, Moscow, ID 83844-3052.

Abbreviations used in this paper: IBMX, 3-isobutyl-1-methylxanthine; IFT, intraflagellar transport; M+M, membrane plus matrix; RSP, radial spoke protein.

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