Site‐directed mutagenesis of Saccharomyces cerevisiae β‐tubulin: interaction between residue 167 and benzimidazole compounds

FEBS Letters

Katiyar, Santosh K.¹  Edlind, Thomas D.¹  Li, Jing¹ 

[1] Department of Microbiology and Immunology, Medical College of Pennsylvania and Hahnemann University, 2900 Queen Lane, Philadelphia, PA 19129, USA

关键词: Benzimidazole;    Microtubule;    β-Tubulin;    Saccharomyces cerevisiae;   

DOI  :  10.1016/0014-5793(96)00334-1

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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.

【 授权许可】

Unknown