A convenient system for the control of gene expression in Saccharomyces cerevisiae was developed. Tetracycline-responsive promoters were constructed by fusing the tetracycline operator (tetO) to the S. cerevisiae HOP1 promoter. When fused to the tetracycline repressor (tetR), trans-activation domains of both GAL4 and HAP4 were capable of promoting transcription from the tetO-HOP1 chimeric promoter, but the tetR-HAP4 fusion activator was the more efficient transcriptional activator. Addition of tetracycline nearly completely repressed activator-dependent transcription from the tetO-HOP1 promoter. Moreover, tetracycline-dependent repression of YEF3, CDC28 and RAM2 expression impaired cell growth. Thus, this system is useful for the elucidation of gene function in S. cerevisiae.