ε-tubulin is essential in Tetrahymena thermophila for the assembly and stability of basal bodies

Ian Ross, Christina Clarissa, Thomas H. Giddings, Mark Winey

Research output: Contribution to journalArticle

19 Scopus citations

Abstract

Basal bodies and centrioles are conserved microtubule-based organelles the improper assembly of which leads to a number of diseases, including ciliopathies and cancer. Tubulin family members are conserved components of these structures that are integral to their proper formation and function. We have identified the ε-tubulin gene in Tetrahymena thermophila and detected the protein, through fluorescence of a tagged allele, to basal bodies. Immunoelectron microscopy has shown that ε-tubulin localizes primarily to the core microtubule scaffold. A complete genomic knockout of ε-tubulin has revealed that it is an essential gene required for the assembly and maintenance of the triplet microtubule blades of basal bodies. We have conducted site-directed mutagenesis of the ε-tubulin gene and shown that residues within the nucleotide-binding domain, longitudinal interacting domains, and C-terminal tail are required for proper function. A single amino acid change of Thr150, a conserved residue in the nucleotide-binding domain, to Val is a conditional mutation that results in defects in the spatial and temporal assembly of basal bodies as well as their stability. We have genetically separated functions for the domains of e-tubulin and identified a novel role for the nucleotide-binding domain in the regulation of basal body assembly and stability.

Original languageEnglish (US)
Pages (from-to)3441-3451
Number of pages11
JournalJournal of Cell Science
Volume126
Issue number15
DOIs
StatePublished - Sep 9 2013
Externally publishedYes

Keywords

  • Basal body
  • Epsilon tubulin
  • Microtubule organizing center
  • Tetrahymena thermophila

ASJC Scopus subject areas

  • Cell Biology

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