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The paclitaxel site in tubulin probed by site‐directed mutagenesis of Saccharomyces cerevisiae β‐tubulin
Author(s) -
Entwistle Ruth A.,
Winefield Robert D.,
Foland Travis B.,
Lushington Gerald H.,
Himes Richard H.
Publication year - 2008
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2008.06.013
Subject(s) - tubulin , paclitaxel , saccharomyces cerevisiae , mutagenesis , directed mutagenesis , biochemistry , yeast , strain (injury) , site directed mutagenesis , mutation , chemistry , biology , microtubule , microbiology and biotechnology , genetics , gene , cancer , mutant , anatomy
Previously, we created a paclitaxel‐sensitive strain of Saccharomyces cerevisiae by mutating five amino acid residues in β‐tubulin in a strain that has a decreased level of the ABC multidrug transporters. We have used site‐directed mutagenesis to examine the relative importance of the five residues in determining sensitivity of this strain to paclitaxel. We found that the change at position 19 from K (brain β‐tubulin) to A (yeast β‐tubulin) and at position 227 from H (brain β‐tubulin) to N (yeast β‐tubulin) had no effect on the activity of paclitaxel. On the other hand, the changes V23T, D26G and F270Y, drastically reduced sensitivity of AD1‐8‐tax to paclitaxel. Molecular modeling and computational studies were used to explain the results.