1887

Abstract

The basidiomycete causes life-threatening infections in immunocompromised patients, and available chemotherapeutic agents are potentially toxic or have limited efficacy. , is very sensitive to selected benzimidazole compounds (e.g. albendazole), which act by disrupting microtubules through binding to the β-tubulin subunit. To understand the basis for this benzimidazole sensitivity, we have characterized β-tubulin genes and their expression. Analysis of PCR amplification products, genomic and cDNA clones and Southern blots identified two β-tubulin genes. contains seven introns, including one that splits the start codon, and encodes a 447 amino acid protein with >80% identity to most other β-tubulins. A partial sequence of revealed a higher density of introns and a considerably more divergent β-tubulin. The relative expression of to determined by reverse-transcription PCR was about 3:1, consistent with a more limited role for the product. Comparisons of β-tubulin sequences from and from various benzimidazole-sensitive and -resistant organisms strongly suggest that the product represents the primary benzimidazole target. This was supported by the identification of a His6 to Gin change in from three independently isolated albendazole-resistant mutants.

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1997-06-01
2021-10-19
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