Abstract
Taxol is a plant alkaloid that has antimitotic activity and appears to stabilize microtubules [Schiff, P. B., Fant, J. & Horwitz, S. B. (1979) Nature (London) 277, 665-667]. Taxol-resistant cells were selected from a population of UV-mutagen-treated Chinese hamster ovary cells by a single-step procedure. These mutants have normal morphologies and growth rates but are 2- to 3-fold more resistant to the toxic effects of the drug than the wild-type parent. One out of 20 mutants screened by two-dimensional electrophoresis for chemical alterations in tubulin had an "extra" spot with a more acidic isoelectric point that alpha-tubulin. This extra spot was shown to be an electrophoretic variant alpha-tubulin by its copurification with tubulin in crude microtubule-containing preparations and by one-dimensional peptide mapping. The alpha-tubulin mutant was found to be temperature sensitive for growth, and this property was used as the basis for the selection of revertants. Seventeen temperature-resistant revertants of the alpha-tubulin mutant were selected for their ability to grow at 40 degrees C and three of these revertants were found to have simultaneously lost their taxol resistance and the electrophoretic variant alpha-tubulin. These results provide evidence that an alteration in alpha-tubulin can confer taxon resistance on a mammalian cell line and suggest that alpha-tubulin is essential for cell viability.
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