Abstract
Azole resistance in Candida albicans can be due to upregulation of multidrug transporters belonging to ABC (ATP-binding cassette) transporters (CDR1 and CDR2) or major facilitators (CaMDR1). Upregulation of these genes can also be achieved by exposure to fluphenazine, resulting in specific upregulation of CDR1 and CDR2 and by exposure to benomyl, resulting in specific CaMDR1 upregulation. In this study, these two different states of gene upregulation were used to determine coregulated genes that often share similar functions or similar regulatory regions. The transcript profiles of a laboratory strain exposed to these drugs were therefore determined and compared with those of two matched pairs of azole-susceptible and -resistant strains expressing CDR1 and CDR2 (CDR strains) or CaMDR1 (MDR isolates). The results obtained revealed that, among 42 commonly regulated genes (8.6% of all regulated genes) between fluphenazine-exposed cells and CDR isolates, the most upregulated were CDR1 and CDR2 as expected, but also IFU5, RTA3 (which encodes putative membrane proteins), HSP12 (which encodes heat shock protein), and IPF4065 (which is potentially involved in stress response). Interestingly, all but HSP12 and IPF4065 contain a putative cis-acting drug responsive element in their promoters. Among the 57 genes (11.5% of all regulated genes) commonly regulated between benomyl-exposed cells and MDR isolates, the most upregulated were CaMDR1 as expected but also genes with oxido-reductive functions such as IFD genes, IPF5987, GRP2 (all belonging to the aldo-keto reductase family), IPF7817 [NAD(P)H oxido-reductase], and IPF17186. Taken together, these results show that in vitro drug-induced gene expression only partially mimics expression profiles observed in azole-resistant clinical strains. Upregulated genes in both drug-exposed conditions and clinical strains are drug resistance genes but also genes that could be activated under cell damage conditions.
MeSH Terms
ATP Binding Cassette Transporter, Subfamily B/biosynthesis,genetics
ATP Binding Cassette Transporter, Subfamily B, Member 1/biosynthesis,genetics
Antifungal Agents/pharmacology
Azoles/pharmacology
Benomyl/pharmacology
Blotting, Northern
Candida albicans/drug effects,genetics
Candidiasis/microbiology
Cluster Analysis
Fluphenazine/pharmacology
Fungal Proteins/biosynthesis,genetics
Fungicides, Industrial/pharmacology
Gene Expression Regulation, Fungal/genetics
Membrane Transport Proteins/biosynthesis,genetics
Oligonucleotide Array Sequence Analysis
RNA, Fungal/biosynthesis,genetics
Regulatory Sequences, Nucleic Acid/genetics
Reproducibility of Results
Reverse Transcriptase Polymerase Chain Reaction
Transcription, Genetic/genetics
Up-Regulation/genetics
Chemicals
ATP Binding Cassette Transporter, Subfamily B
ATP Binding Cassette Transporter, Subfamily B, Member 1
Antifungal Agents
Azoles
CDR1 protein, Candida albicans
Fungal Proteins
Fungicides, Industrial
Membrane Transport Proteins
RNA, Fungal
Fluphenazine
Benomyl
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Karababa Mahir
Institut de Microbiologie, Centre Hospitalier Universitaire Vaudois, 1011 Lausanne, Switzerland.
Coste Alix T
Rognon Bénédicte
Bille Jacques
Sanglard Dominique
References (33)
33 references, click to expand
-
Population genomics of drug resistance in Candida albicans.
Proc Natl Acad Sci U S A. 2002 Jul 9;99(14):9284-9
PMID: 12089321
-
A common drug-responsive element mediates the upregulation of the Candida albicans ABC transporters CDR1 and CDR2, two genes involved in antifungal drug resistance.
Mol Microbiol. 2002 Mar;43(5):1197-214
PMID: 11918807
-
Genome-wide expression profile analysis reveals coordinately regulated genes associated with stepwise acquisition of azole resistance in Candida albicans clinical isolates.
Antimicrob Agents Chemother. 2003 Apr;47(4):1220-7
PMID: 12654650
-
Stress-induced gene expression in Candida albicans: absence of a general stress response.
Mol Biol Cell. 2003 Apr;14(4):1460-7
PMID: 12686601
-
Genome-wide expression profiling of the response to polyene, pyrimidine, azole, and echinocandin antifungal agents in Saccharomyces cerevisiae.
J Biol Chem. 2003 Sep 12;278(37):34998-5015
PMID: 12824174
-
"A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity". Addendum.
Anal Biochem. 1984 Feb;137(1):266-7
PMID: 6329026
-
Isogenic strain construction and gene mapping in Candida albicans.
Genetics. 1993 Jul;134(3):717-28
PMID: 8349105
-
Expression of seven members of the gene family encoding secretory aspartyl proteinases in Candida albicans.
Mol Microbiol. 1994 Oct;14(1):87-99
PMID: 7830564
-
Resistance of Candida species to fluconazole.
Antimicrob Agents Chemother. 1995 Jan;39(1):1-8
PMID: 7695288
-
The ATP binding cassette transporter gene CgCDR1 from Candida glabrata is involved in the resistance of clinical isolates to azole antifungal agents.
Antimicrob Agents Chemother. 1999 Nov;43(11):2753-65
PMID: 10543759
-
Activation of the multiple drug resistance gene MDR1 in fluconazole-resistant, clinical Candida albicans strains is caused by mutations in a trans-regulatory factor.
J Bacteriol. 2000 Jan;182(2):400-4
PMID: 10629186
-
Evolution of drug resistance in experimental populations of Candida albicans.
J Bacteriol. 2000 Mar;182(6):1515-22
PMID: 10692355
-
Genome microarray analysis of transcriptional activation in multidrug resistance yeast mutants.
FEBS Lett. 2000 Mar 24;470(2):156-60
PMID: 10734226
-
Genome-wide expression patterns in Saccharomyces cerevisiae: comparison of drug treatments and genetic alterations affecting biosynthesis of ergosterol.
Antimicrob Agents Chemother. 2000 May;44(5):1255-65
PMID: 10770760
-
Analysis of the oxidative stress regulation of the Candida albicans transcription factor, Cap1p.
Mol Microbiol. 2000 May;36(3):618-29
PMID: 10844651
-
A novel multidrug efflux transporter gene of the major facilitator superfamily from Candida albicans (FLU1) conferring resistance to fluconazole.
Microbiology. 2000 Nov;146 ( Pt 11):2743-54
PMID: 11065353
-
Role of ATP-binding-cassette transporter genes in high-frequency acquisition of resistance to azole antifungals in Candida glabrata.
Antimicrob Agents Chemother. 2001 Apr;45(4):1174-83
PMID: 11257032
-
Genomic profiling of the response of Candida albicans to itraconazole treatment using a DNA microarray.
Antimicrob Agents Chemother. 2001 Jun;45(6):1660-70
PMID: 11353609
-
Mechanisms of resistance to azole antifungal agents in Candida albicans isolates from AIDS patients involve specific multidrug transporters.
Antimicrob Agents Chemother. 1995 Nov;39(11):2378-86
PMID: 8585712
-
Typing Candida albicans oral isolates from human immunodeficiency virus-infected patients by multilocus enzyme electrophoresis and DNA fingerprinting.
J Clin Microbiol. 1996 May;34(5):1235-48
PMID: 8727910
-
The changing face of candidemia: emergence of non-Candida albicans species and antifungal resistance.
Am J Med. 1996 Jun;100(6):617-23
PMID: 8678081
-
Cloning of Candida albicans genes conferring resistance to azole antifungal agents: characterization of CDR2, a new multidrug ABC transporter gene.
Microbiology. 1997 Feb;143 ( Pt 2):405-16
PMID: 9043118
-
Increased mRNA levels of ERG16, CDR, and MDR1 correlate with increases in azole resistance in Candida albicans isolates from a patient infected with human immunodeficiency virus.
Antimicrob Agents Chemother. 1997 Jul;41(7):1482-7
PMID: 9210670
-
The presence of an R467K amino acid substitution and loss of allelic variation correlate with an azole-resistant lanosterol 14alpha demethylase in Candida albicans.
Antimicrob Agents Chemother. 1997 Jul;41(7):1488-94
PMID: 9210671
-
Yap, a novel family of eight bZIP proteins in Saccharomyces cerevisiae with distinct biological functions.
Mol Cell Biol. 1997 Dec;17(12):6982-93
PMID: 9372930
-
Amino acid substitutions in the cytochrome P-450 lanosterol 14alpha-demethylase (CYP51A1) from azole-resistant Candida albicans clinical isolates contribute to resistance to azole antifungal agents.
Antimicrob Agents Chemother. 1998 Feb;42(2):241-53
PMID: 9527767
-
Identification of polymorphic mutant alleles of CaMDR1, a major facilitator of Candida albicans which confers multidrug resistance, and its in vitro transcriptional activation.
Curr Genet. 1998 Sep;34(3):192-9
PMID: 9745021
-
Cluster analysis and display of genome-wide expression patterns.
Proc Natl Acad Sci U S A. 1998 Dec 8;95(25):14863-8
PMID: 9843981
-
The bZip transcription factor Cap1p is involved in multidrug resistance and oxidative stress response in Candida albicans.
J Bacteriol. 1999 Feb;181(3):700-8
PMID: 9922230
-
Cloning and analysis of a Candida albicans gene that affects cell surface hydrophobicity.
J Bacteriol. 2001 Jun;183(12):3582-8
PMID: 11371521
-
Regulation of the yeast Yap1p nuclear export signal is mediated by redox signal-induced reversible disulfide bond formation.
Mol Cell Biol. 2001 Sep;21(18):6139-50
PMID: 11509657
-
Prevalence of molecular mechanisms of resistance to azole antifungal agents in Candida albicans strains displaying high-level fluconazole resistance isolated from human immunodeficiency virus-infected patients.
Antimicrob Agents Chemother. 2001 Oct;45(10):2676-84
PMID: 11557454
-
Genome-wide analysis of gene expression regulated by the calcineurin/Crz1p signaling pathway in Saccharomyces cerevisiae.
J Biol Chem. 2002 Aug 23;277(34):31079-88
PMID: 12058033