Category MI L20 Multidrug Resistance: C. albicans Gene IPT1 Affects Gene

Expression

Abstract Candida albicans is the most commonly isolated yeast in human disease; it

acutely impacts immunocompromised patients. Furthermore, C. albicans

resistance to antifungal drugs is escalating. The purpose of this

investigation was to determine which C. albicans genes in the target

pathways of the antifungal drugs: aureobasidin, rapamycin, and

fluvastatin are affected by transformation with high copy number plasmids

containing the gene IPT1. The two transformants and the controls were

cultured and tested on ten antifungal drugs at five concentrations, and the

transformants were proven resistant to nine of these drugs. RNA was

isolated from the transformants at 4 time intervals with exposure to three

drugs and no drug. Total mRNA was reverse transcribed into cDNA,

which was then quantified by real-time PCR. Levels of transcription of IPT1

and of genes known to affect drug resistance were determined, including

AUR1 (aureobasidin A), HMG1 (fluvastatin), and TOR (rapamycin), and

known drug efflux genes (CDR1 and MDR1). Data indicated that

expression of the cloned IPT1 was not necessary for resistance,

suggesting that DNA sequences are directly altering phenotype, perhaps

by sequestering regulatory proteins. Genomic DNA agarose gel analysis

revealed that one plasmid had a rearranged version of IPT1 and both

plasmids had an insert. The observed changes in expression likely cause

the multidrug resistance and occur because of integrative disruption of

chromosomal IPT1 by the homologous sequences in the plasmid. Multidrug

resistance suggests that there are regulatory links between unrelated

pathways that can be utilized for drug development.

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