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© The Rockefeller University Press, 0021-9525/1998//1591 $5.00
The Journal of Cell Biology, Volume 143, Number 6, , 1998 1591-1601

Distinct Mutants of Retrograde Intraflagellar Transport (IFT) Share Similar Morphological and Molecular Defects

Gianni Piperno^*, Edward Siuda^*, Scott Henderson^*, Margarethe Segil^*, Heikki Vaananen, and Massimo Sassaroli

^* Department of Cell Biology and Anatomy and Department of Physiology and Biophysics, Mount Sinai School of Medicine, New York, 10029

A microtubule-based transport of protein complexes, which is bidirectional and occurs between the space surrounding the basal bodies and the distal part of Chlamydomonas flagella, is referred to as intraflagellar transport (IFT). The IFT involves molecular motors and particles that consist of 17S protein complexes. To identify the function of different components of the IFT machinery, we isolated and characterized four temperature-sensitive (ts) mutants of flagellar assembly that represent the loci FLA15, FLA16, and FLA17. These mutants were selected among other ts mutants of flagellar assembly because they displayed a characteristic bulge of the flagellar membrane as a nonconditional phenotype. Each of these mutants was significantly defective for the retrograde velocity of particles and the frequency of bidirectional transport but not for the anterograde velocity of particles, as revealed by a novel method of analysis of IFT that allows tracking of single particles in a sequence of video images. Furthermore, each mutant was defective for the same four subunits of a 17S complex that was identified earlier as the IFT complex A. The occurrence of the same set of phenotypes, as the result of a mutation in any one of three loci, suggests the hypothesis that complex A is a portion of the IFT particles specifically involved in retrograde intraflagellar movement.

Key Words: Chlamydomonas • temperature-sensitive mutants • intraflagellar particles • video microscopy • retrograde transport

Abbreviations used in this paper: IFT, intraflagellar transport; MNNG, N-methyl-N'-nitro-N nitrosoguanidine; ts, temperature-sensitive.

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