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 isolated from other systems. The gene was adapted for expression in E. coli and yielded a soluble and highly-active enzyme which is easily purified. 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. cerevisiae, and deletion of the MluI site resulted in a large reduction of DUT1 transcription in C. albicans. These results suggest that MCB elements 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.
McIntosh
E M
EM
Department of Biology, University of York, Toronto, Ontario, Canada.
Looser
J
J
Haynes
R H
RH
Pearlman
R E
RE
D000595:Amino Acid Sequence; D001426:Bacterial Proteins; D001483:Base Sequence; D015152:Blotting, Northern; D002176:Candida albicans / Q000201:enzymology* / Q000235:genetics*; D003001:Cloning, Molecular; D017931:DNA Primers; D004261:DNA Replication; D015252:Deoxyribonucleases, Type II Site-Specific; D004926:Escherichia coli / Q000235:genetics; D015971:Gene Expression Regulation, Enzymologic; D015966:Gene Expression Regulation, Fungal; D005800:Genes, Fungal; D006801:Humans; D016089:Immunodeficiency Virus, Feline / Q000235:genetics; D018551:Solanum lycopersicum / Q000235:genetics; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011755:Pyrophosphatases / Q000096:biosynthesis* / Q000235:genetics; D012331:RNA, Fungal / Q000032:analysis / Q000096:biosynthesis; D011994:Recombinant Proteins / Q000096:biosynthesis; D015183:Restriction Mapping; D017386:Sequence Homology, Amino Acid; D014158:Transcription, Genetic
D001426:Bacterial Proteins; D017931:DNA Primers; D012331:RNA, Fungal; D011994:Recombinant Proteins; C035041:endodeoxyribonuclease MluI; D015252:Deoxyribonucleases, Type II Site-Specific; D011755:Pyrophosphatases; C020020:dUTP pyrophosphatase