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© The Rockefeller University Press, 0021-9525/1998//993 $5.00
The Journal of Cell Biology, Volume 141, Number 4, , 1998 993-1008

Articles

Chlamydomonas Kinesin-II–dependent Intraflagellar Transport (IFT): IFT Particles Contain Proteins Required for Ciliary Assembly in Caenorhabditis elegans Sensory Neurons



Douglas G. Cole*, Dennis R. Diener*, Amy L. Himelblau*, Peter L. Beech{ddagger}, Jason C. Fuster*, and Joel L. Rosenbaum*

* Molecular, Cellular and Developmental Biology, Yale University, New Haven, Connecticut 06520-8103; {ddagger} School of Botany, University of Melbourne, Parkville, 3052, Victoria, Australia

We previously described a kinesin-dependent movement of particles in the flagella of Chlamydomonas reinhardtii called intraflagellar transport (IFT) (Kozminski, K.G., K.A. Johnson, P. Forscher, and J.L. Rosenbaum. 1993. Proc. Natl. Acad. Sci. USA. 90:5519–5523). When IFT is inhibited by inactivation of a kinesin, FLA10, in the temperature-sensitive mutant, fla10, existing flagella resorb and new flagella cannot be assembled. We report here that: (a) the IFT-associated FLA10 protein is a subunit of a heterotrimeric kinesin; (b) IFT particles are composed of 15 polypeptides comprising two large complexes; (c) the FLA10 kinesin-II and IFT particle polypeptides, in addition to being found in flagella, are highly concentrated around the flagellar basal bodies; and, (d) mutations affecting homologs of two of the IFT particle polypeptides in Caenorhabditis elegans result in defects in the sensory cilia located on the dendritic processes of sensory neurons. In the accompanying report by Pazour, G.J., C.G. Wilkerson, and G.B. Witman (1998. J. Cell Biol. 141:979–992), a Chlamydomonas mutant (fla14) is described in which only the retrograde transport of IFT particles is disrupted, resulting in assembly-defective flagella filled with an excess of IFT particles. This microtubule- dependent transport process, IFT, defined by mutants in both the anterograde (fla10) and retrograde (fla14) transport of isolable particles, is probably essential for the maintenance and assembly of all eukaryotic motile flagella and nonmotile sensory cilia.

Abbreviations used in this paper: AMPPNP, 5'-adenylylimidodiphosphate; FLA10, the fla10 gene product; FLA10 kinesin-II, heterotrimeric complex containing FLA10 and two additional subunits; IFT, intraflagellar transport; MT, microtubule; ts, temperature-sensitive.

Address all correspondence to Joel Rosenbaum, MCDB KBT 310, Yale University, P.O. Box 208103, New Haven, CT 06520-8103. Tel.: (203) 432-3472. Fax: (203) 432-5059. E-mail: joel.rosenbaum{at}yale.edu

Brief reports of this work were presented at the International Congress on Chlamydomonas in Regensburg, Germany on May 27–June 1, 1996, and appeared in abstract form (Cole, D.G., and J.L. Rosenbaum. 1996. Mol. Biol. Cell. 7S:47a; Cole, D.G., D.R. Diener, J. Fuster, A. Himelblau, and J.L. Rosenbaum. 1997. Mol. Biol. Cell. 8S:54a).


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