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Published online 2 April 2001. doi:10.1083/jcb.153.1.13
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© The Rockefeller University Press, 0021-9525/2001//13 $5.00
The Journal of Cell Biology, Volume 153, Number 1, , 2001 13-24

Original Article

Protein Particles in Chlamydomonas Flagella Undergo a Transport Cycle Consisting of Four Phases



Carlo Iominia, Veronica Babaev-Khaimova, Massimo Sassarolib, and Gianni Pipernoa

a Department of Cell Biology and Anatomy, Mount Sinai School of Medicine, New York, New York 10029
b Department of Physiology and Biophysics, Mount Sinai School of Medicine, New York, New York 10029
Department of Cell Biology and Anatomy, Mount Sinai School of Medicine, 1 Gustave L. Levy Pl., Box 1007, New York, NY 10029.(212) 860-1174(212) 241-0773

piperno{at}msvax.mssm.edu

We used an improved procedure to analyze the intraflagellar transport (IFT) of protein particles in Chlamydomonas and found that the frequency of the particles, not only the velocity, changes at each end of the flagella. Thus, particles undergo structural remodeling at both flagellar locations. Therefore, we propose that the IFT consists of a cycle composed of at least four phases: phases II and IV, in which particles undergo anterograde and retrograde transport, respectively, and phases I and III, in which particles are remodeled/exchanged at the proximal and distal end of the flagellum, respectively. In support of our model, we also identified 13 distinct mutants of flagellar assembly (fla), each defective in one or two consecutive phases of the IFT cycle. The phase I-II mutant fla10-1 revealed that cytoplasmic dynein requires the function of kinesin II to participate in the cycle. Phase I and II mutants accumulate complex A, a particle component, near the basal bodies. In contrast, phase III and IV mutants accumulate complex B, a second particle component, in flagellar bulges. Thus, fla mutations affect the function of each complex at different phases of the cycle.

Key Words: intraflagellar transport • transport cycle • Chlamydomonas • temperature-sensitive mutants • kinesin II

© 2001 The Rockefeller University Press

Abbreviations used in this paper: Ab, antibody; DHC, dynein heavy chain; IFT, intraflagellar transport; KHP, kinesin homologue protein; ts, temperature-sensitive; VE-DIC, video-enhanced differential interference contrast.


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