The inducible N-acetylglucosamine catabolic pathway gene cluster in Candida albicans: Discrete N-acetylglucosamine-inducible factors interact at the promoter of NAG1

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The National Academy of Sciences

RESUMO

The catabolic pathway of N-acetylglucosamine (GlcNAc) in Candida albicans is an important facet of its pathogenicity. One of the pathway genes, encoding glucosamine-6-phosphate deaminase (NAG1) is transcriptionally regulated by GlcNAc. Sequence analysis of a 4-kb genomic clone containing NAG1 indicates that this gene is part of a cluster containing two other genes of the GlcNAc catabolic pathway, i.e., DAC1, GlcNAc-6-phosphate deacetylase, and HXK1, hexokinase. All three genes are temporally and coordinately induced by GlcNAc suggesting a common regulatory mechanism for these genes. The NAG1 promoter is up-regulated when induced by GlcNAc in C. albicans but not in Saccharomyces cerevisiae. In vivo analysis of the deletion constructs delineated the minimal promoter to −130 bp and mapped two regions at −200 and −400 bp upstream of +1 (ATG) responsible for GlcNAc induction. Gel mobility-shift assays and “footprinting” (DNase protection method) analyses revealed two regions, 5′-GGAGCAAAAAAATGT 3′ (−164 to −150, box A) and 5′-ACGGTGAGTTG 3′ (−291 to −281, box B), that are recognized and bound by at least two inducible activator proteins directing the regulation of gene expression.

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