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Site‐directed mutagenesis of Saccharomyces cerevisiae β‐tubulin: interaction between residue 167 and benzimidazole compounds
Author(s) -
Li Jing,
Katiyar Santosh K.,
Edlind Thomas D.
Publication year - 1996
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 - 10.1016/0014-5793(96)00334-1
Subject(s) - benzimidazole , benomyl , saccharomyces cerevisiae , mutant , mutagenesis , biochemistry , site directed mutagenesis , chemistry , carbendazim , nocodazole , tubulin , microtubule , residue (chemistry) , directed mutagenesis , biology , stereochemistry , yeast , gene , fungicide , microbiology and biotechnology , botany , cell , organic chemistry , cytoskeleton
Benzimidazoles are widely used as anthelmintic agents and systemic fungicides. In susceptible organisms, benzimidazoles bind to β‐tubulin and block microtubule polymerization. To further characterize this interaction, site‐directed mutagenesis followed by gene replacement was used to change Saccharomyces cerevisiae β‐tubulin residue Phe‐167 to Tyr. Consistent with previous studies, this mutation resulted in at least 3–4‐fold decreased sensitivity to the benzimidazole derivatives carbendazim and nocodazole. The Tyr‐167 mutant was cold sensitive, implying a direct effect on benzimidazole binding rather than a nonspecific increase in microtubule stability. Surprisingly, the mutant had 8‐fold increased sensitivity to the derivative benomyl, which is structurally identical to carbendazim except at position 1. This suggests that residue 167 interacts with benzimidazoles in the vicinity of the 1‐position.