Original Article

Polyglycylation of Tubulin Is Essential and Affects Cell Motility and Division in Tetrahymena thermophila

Correspondence to: Jacek Gaertig, Department of Cellular Biology, 724 Biological Sciences Building, University of Georgia, Athens, GA 30602-2607. Tel:(706) 542-3409 Fax:(706) 542-4271 E-mail:jgaertig{at}cb.uga.edu.

We analyzed the role of tubulin polyglycylation in Tetrahymena thermophila using in vivo mutagenesis and immunochemical analysis with modification-specific antibodies. Three and five polyglycylation sites were identified at glutamic acids near the COOH termini of - and ß-tubulin, respectively. Mutants lacking all polyglycylation sites on -tubulin have normal phenotype, whereas similar sites on ß-tubulin are essential. A viable mutant with three mutated sites in ß-tubulin showed reduced tubulin glycylation, slow growth and motility, and defects in cytokinesis. Cells in which all five polyglycylation sites on ß-tubulin were mutated were viable if they were cotransformed with an -tubulin gene whose COOH terminus was replaced by the wild-type COOH terminus of ß-tubulin. In this double mutant, ß-tubulin lacked detectable polyglycylation, while the -ß tubulin chimera was hyperglycylated compared with -tubulin in wild-type cells. Thus, the essential function of polyglycylation of the COOH terminus of ß-tubulin can be transferred to -tubulin, indicating it is the total amount of polyglycylation on both - and ß-tubulin that is essential for survival.

Key Words: motor proteins, microtubules, cilia, cytoskeleton, motility

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