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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 α- and β-tubulin, respectively. Muta...

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Autores principales: Xia, Lu, Hai, Bing, Gao, Yan, Burnette, Dylan, Thazhath, Rupal, Duan, Jianming, Bré, Marie-Helene, Levilliers, Nicolette, Gorovsky, Martin A., Gaertig, Jacek
Formato: Texto
Lenguaje:English
Publicado: The Rockefeller University Press 2000
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2174830/
https://www.ncbi.nlm.nih.gov/pubmed/10831613
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author Xia, Lu
Hai, Bing
Gao, Yan
Burnette, Dylan
Thazhath, Rupal
Duan, Jianming
Bré, Marie-Helene
Levilliers, Nicolette
Gorovsky, Martin A.
Gaertig, Jacek
author_facet Xia, Lu
Hai, Bing
Gao, Yan
Burnette, Dylan
Thazhath, Rupal
Duan, Jianming
Bré, Marie-Helene
Levilliers, Nicolette
Gorovsky, Martin A.
Gaertig, Jacek
author_sort Xia, Lu
collection PubMed
description 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 α- and β-tubulin, respectively. Mutants lacking all polyglycylation sites on α-tubulin have normal phenotype, whereas similar sites on β-tubulin are essential. A viable mutant with three mutated sites in β-tubulin showed reduced tubulin glycylation, slow growth and motility, and defects in cytokinesis. Cells in which all five polyglycylation sites on β-tubulin were mutated were viable if they were cotransformed with an α-tubulin gene whose COOH terminus was replaced by the wild-type COOH terminus of β-tubulin. In this double mutant, β-tubulin lacked detectable polyglycylation, while the α-β tubulin chimera was hyperglycylated compared with α-tubulin in wild-type cells. Thus, the essential function of polyglycylation of the COOH terminus of β-tubulin can be transferred to α-tubulin, indicating it is the total amount of polyglycylation on both α- and β-tubulin that is essential for survival.
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spelling pubmed-21748302008-05-01 Polyglycylation of Tubulin Is Essential and Affects Cell Motility and Division in Tetrahymena thermophila Xia, Lu Hai, Bing Gao, Yan Burnette, Dylan Thazhath, Rupal Duan, Jianming Bré, Marie-Helene Levilliers, Nicolette Gorovsky, Martin A. Gaertig, Jacek J Cell Biol Original Article 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 α- and β-tubulin, respectively. Mutants lacking all polyglycylation sites on α-tubulin have normal phenotype, whereas similar sites on β-tubulin are essential. A viable mutant with three mutated sites in β-tubulin showed reduced tubulin glycylation, slow growth and motility, and defects in cytokinesis. Cells in which all five polyglycylation sites on β-tubulin were mutated were viable if they were cotransformed with an α-tubulin gene whose COOH terminus was replaced by the wild-type COOH terminus of β-tubulin. In this double mutant, β-tubulin lacked detectable polyglycylation, while the α-β tubulin chimera was hyperglycylated compared with α-tubulin in wild-type cells. Thus, the essential function of polyglycylation of the COOH terminus of β-tubulin can be transferred to α-tubulin, indicating it is the total amount of polyglycylation on both α- and β-tubulin that is essential for survival. The Rockefeller University Press 2000-05-29 /pmc/articles/PMC2174830/ /pubmed/10831613 Text en © 2000 The Rockefeller University Press This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).
spellingShingle Original Article
Xia, Lu
Hai, Bing
Gao, Yan
Burnette, Dylan
Thazhath, Rupal
Duan, Jianming
Bré, Marie-Helene
Levilliers, Nicolette
Gorovsky, Martin A.
Gaertig, Jacek
Polyglycylation of Tubulin Is Essential and Affects Cell Motility and Division in Tetrahymena thermophila
title Polyglycylation of Tubulin Is Essential and Affects Cell Motility and Division in Tetrahymena thermophila
title_full Polyglycylation of Tubulin Is Essential and Affects Cell Motility and Division in Tetrahymena thermophila
title_fullStr Polyglycylation of Tubulin Is Essential and Affects Cell Motility and Division in Tetrahymena thermophila
title_full_unstemmed Polyglycylation of Tubulin Is Essential and Affects Cell Motility and Division in Tetrahymena thermophila
title_short Polyglycylation of Tubulin Is Essential and Affects Cell Motility and Division in Tetrahymena thermophila
title_sort polyglycylation of tubulin is essential and affects cell motility and division in tetrahymena thermophila
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2174830/
https://www.ncbi.nlm.nih.gov/pubmed/10831613
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