MLUI SITE-DEPENDENT TRANSCRIPTIONAL REGULATION OF THE CANDIDA-ALBICANS DUTPASE GENE

The Candida albicans dUTP pyrophosphatase (dUTPase) gene DUT1 has been isolated by genetic complementation in S. cerevisiae. it was found to encode a 17-kDa protein similar in amino-acid sequence to dUTPases is olated from other systems. The gene was adapted for expression in E. c oli and yielded a soluble and highly-active enzyme which is easily pur ified. The 5' flanking sequence of DUT1 contains an MluI site typical of MCB cell-cycle-dependent UAS elements of budding and fission yeast. We found the gene to be cell-cycle-regulated when expressed in S. cer evisiae, and deletion of the MlUI site resulted in a large reduction o f DUT1 transcription in C. albicans. These results suggest that MCB el ements are functionally conserved in this pathogenic fungus. Based on the vital role that dUTPase plays in DNA replication, the C. albicans enzyme may be a potentially useful target for the development of novel anti-fungal compounds.