Identification and characterization of Cor33p, a novel protein implicated in tolerance towards oxidative stress in Candida albicans.
We applied two-dimensional gel electrophoresis to identify downstream effectors of CPH1 and EFG1 under hypha-inducing conditions in Candida albicans. Among the proteins that were expressed in wild-type cells but were strongly downregulated in a cph1Delta/efg1Delta double mutant in alpha-minimal essential medium at 37 degrees C, we could identify not-yet-characterized proteins, including Cor33-1p and Cor33-2p. The two proteins are almost identical (97% identity) and represent products of allelic isoforms of the same gene. Cor33p is highly similar to Cip1p from Candida sp. but lacks any significant homology to proteins from Saccharomyces cerevisiae. Strikingly, both proteins share homology with phenylcoumaran benzylic ether reductases and isoflavone reductases from plants. For other hypha-inducing media, like yeast-peptone-dextrose (YPD) plus serum at 37 degrees C, we could not detect any transcription for COR33 in wild-type cells, indicating that Cor33p is not hypha specific. In contrast, we found a strong induction for COR33 when cells were treated with 5 mM hydrogen peroxide. However, under oxidative conditions, transcription of COR33 was not dependent on EFG1, indicating that other regulatory factors are involved. In fact, upregulation depends on CAP1 at least, as transcript levels were clearly reduced in a Deltacap1 mutant strain under oxidative conditions. Unlike in wild-type cells, transcription of COR33 in a tsa1Delta mutant can be induced by treatment with 0.1 mM hydrogen peroxide. This suggests a functional link between COR33 and thiol-specific antioxidant-like proteins that are important in the oxidative-stress response in yeasts. Concordantly, cor33Delta deletion mutants show retarded growth on YPD plates supplemented with hydrogen peroxide, indicating that COR33 in general is implicated in conferring tolerance toward oxidative stress on Candida albicans.
Sohn
K
K
Fraunhofer, IGP, Inst. f. Grenzflächen- und Bioverfahrenstechnik, Nobelstr. 12, 70569 Stuttgart, Germany.
Roehm
M
M
Urban
C
C
Saunders
N
N
Rothenstein
D
D
Lottspeich
F
F
Schröppel
K
K
Brunner
H
H
Rupp
S
S
D000483:Alleles; D000595:Amino Acid Sequence; D002176:Candida albicans / Q000737:chemistry* / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D002457:Cell Extracts / Q000737:chemistry; D015825:Chromosomes, Fungal / Q000737:chemistry; D004271:DNA, Fungal / Q000737:chemistry / Q000302:isolation & purification; D030541:Databases, Genetic; D015536:Down-Regulation; D015180:Electrophoresis, Gel, Two-Dimensional; D005656:Fungal Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D017353:Gene Deletion; D015966:Gene Expression Regulation, Fungal; D005800:Genes, Fungal; D006360:Heat-Shock Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D006861:Hydrogen Peroxide / Q000494:pharmacology; D008969:Molecular Sequence Data; D016877:Oxidants / Q000494:pharmacology; D018384:Oxidative Stress; D020033:Protein Isoforms / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D012331:RNA, Fungal / Q000737:chemistry / Q000302:isolation & purification; D017386:Sequence Homology, Amino Acid; D014158:Transcription, Genetic / Q000187:drug effects; D015854:Up-Regulation
D002457:Cell Extracts; D004271:DNA, Fungal; D005656:Fungal Proteins; D006360:Heat-Shock Proteins; D016877:Oxidants; D020033:Protein Isoforms; D012331:RNA, Fungal; D006861:Hydrogen Peroxide