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© The Rockefeller University Press, 0021-9525/1998//979 $5.00
The Journal of Cell Biology, Volume 141, Number 4, , 1998 979-992

Articles

A Dynein Light Chain Is Essential for the Retrograde Particle Movement of Intraflagellar Transport (IFT)



Gregory J. Pazour, Curtis G. Wilkerson, and George B. Witman

Department of Cell Biology, University of Massachusetts Medical Center (UMMC), Worcester Foundation Campus, Shrewsbury, Massachusetts 01545

Several enzymes, including cytoplasmic and flagellar outer arm dynein, share an Mr 8,000 light chain termed LC8. The function of this chain is unknown, but it is highly conserved between a wide variety of organisms. We have identified deletion alleles of the gene (fla14) encoding this protein in Chlamydomonas reinhardtii. These mutants have short, immotile flagella with deficiencies in radial spokes, in the inner and outer arms, and in the beak-like projections in the B tubule of the outer doublet microtubules. Most dramatically, the space between the doublet microtubules and the flagellar membrane contains an unusually high number of rafts, the particles translocated by intraflagellar transport (IFT) (Kozminski, K.G., P.L. Beech, and J.L. Rosenbaum. 1995. J. Cell Biol. 131:1517–1527). IFT is a rapid bidirectional movement of rafts under the flagellar membrane along axonemal microtubules. Anterograde IFT is dependent on a kinesin whereas the motor for retrograde IFT is unknown. Anterograde IFT is normal in the LC8 mutants but retrograde IFT is absent; this undoubtedly accounts for the accumulation of rafts in the flagellum. This is the first mutation shown to specifically affect retrograde IFT; the fact that LC8 loss affects retrograde IFT strongly suggests that cytoplasmic dynein is the motor that drives this process. Concomitant with the accumulation of rafts, LC8 mutants accumulate proteins that are components of the 15-16S IFT complexes (Cole, D.G., D.R. Deiner, A.L. Himelblau, P.L. Beech, J.C. Fuster, and J.L. Rosenbaum. 1998. J. Cell Biol. 141:993–1008), confirming that these complexes are subunits of the rafts. Polystyrene microbeads are still translocated on the surface of the flagella of LC8 mutants, indicating that the motor for flagellar surface motility is different than the motor for retrograde IFT.

Abbreviations used in this paper: DHC, dynein heavy chain; DIC, differential interference contrast; IFT, intraflagellar transport.

Address all correspondence to George Witman, Department of Cell Biology, UMMC, Worcester Foundation Campus, 222 Maple Ave., Shrewsbury, MA 01545. Tel.: (508) 842-8921 Ext. 344. Fax: (508) 842-3915. E-mail: witman{at}sci.wfbr.edu

C. Wilkerson's present address is Michigan State University, Department of Energy Plant Research Laboratory and Department of Botany and Plant Pathology, Michigan State University, East Lansing, MI 48824-1312.


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