Polyglycylation of tubulin is essential and affects cell motility and division in Tetrahymena thermophila.
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 alpha- and beta-tubulin, respectively. Mutants lacking all polyglycylation sites on alpha-tubulin have normal phenotype, whereas similar sites on beta-tubulin are essential. A viable mutant with three mutated sites in beta-tubulin showed reduced tubulin glycylation, slow growth and motility, and defects in cytokinesis. Cells in which all five polyglycylation sites on beta-tubulin were mutated were viable if they were cotransformed with an alpha-tubulin gene whose COOH terminus was replaced by the wild-type COOH terminus of beta-tubulin. In this double mutant, beta-tubulin lacked detectable polyglycylation, while the alpha-beta tubulin chimera was hyperglycylated compared with alpha-tubulin in wild-type cells. Thus, the essential function of polyglycylation of the COOH terminus of beta-tubulin can be transferred to alpha-tubulin, indicating it is the total amount of polyglycylation on both alpha- and beta-tubulin that is essential for survival.
Xia
L
L
Department of Cellular Biology, University of Georgia, Athens, Georgia 30602-2607, USA.
Hai
B
B
Gao
Y
Y
Burnette
D
D
Thazhath
R
R
Duan
J
J
Bré
M H
MH
Levilliers
N
N
Gorovsky
M A
MA
Gaertig
J
J
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D002455:Cell Division / Q000502:physiology; D002465:Cell Movement / Q000502:physiology*; D002470:Cell Survival / Q000502:physiology; D002923:Cilia / Q000502:physiology; D006031:Glycosylation; D018613:Microscopy, Confocal; D008870:Microtubules / Q000378:metabolism; D020409:Molecular Motor Proteins / Q000378:metabolism; D008969:Molecular Sequence Data; D016296:Mutagenesis / Q000502:physiology; D010641:Phenotype; D016808:Tetrahymena thermophila / Q000166:cytology* / Q000235:genetics / Q000378:metabolism; D014404:Tubulin / Q000235:genetics* / Q000276:immunology / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D020409:Molecular Motor Proteins; D014404:Tubulin