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Vol. 14, Issue 7, 2999-3012, July 2003

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Three-dimensional Organization of Basal Bodies from Wild-Type and -Tubulin Deletion Strains of Chlamydomonas reinhardtii

Eileen T. O'Toole ^* , Thomas H. Giddings ^*, J. Richard McIntosh ^*, and Susan K. Dutcher 

^* Boulder Laboratory for 3-D Fine Structure, Department of Molecular, Cellular, and Developmental Biology, University of Colorado, Boulder, CO 80309-0347; Department of Genetics, Washington University School of Medicine, St. Louis, Missouri 63110

Submitted November 21, 2002; Revised January 30, 2003; Accepted February 12, 2003
Monitoring Editor: Mary Beckerle

Improved methods of specimen preparation and dual-axis electron tomography have been used to study the structure and organization of basal bodies in the unicellular alga Chlamydomonas reinhardtii. Novel structures have been found in both wild type and strains with mutations that affect specific tubulin isoforms. Previous studies have shown that strains lacking -tubulin fail to assemble the C-tubule of the basal body. Tomographic reconstructions of basal bodies from the -tubulin deletion mutant uni3-1 have confirmed that basal bodies contain mostly doublet microtubules. Our methods now show that the stellate fibers, which are present only in the transition zone of wild-type cells, repeat within the core of uni3-1 basal bodies. The distal striated fiber is incomplete in this mutant, rootlet microtubules can be misplaced, and multiflagellate cells have been observed. A suppressor of uni3-1, designated tua2-6, contains a mutation in -tubulin. tua2-6; uni3-1 cells build both flagella, yet they retain defects in basal body structure and in rootlet microtubule positioning. These data suggest that the presence of specific tubulin isoforms in Chlamydomonas directly affects the assembly and function of both basal bodies and basal body-associated structures.

Corresponding author. E-mail address: eileen{at}bio3d.colorado.edu.

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