Isolation of a Candida albicans gene, tightly linked to URA3, coding for a putative transcription factor that suppresses a Saccharomyces cerevisiae aft1 mutation.
A pathogen such as C. albicans needs an efficient mechanism of iron uptake in an iron-restricted environment such as is the human body. A ferric-reductase activity regulated by iron and copper, and analogous to that in S. cerevisiae, has been described in C. albicans. We have developed an in-plate protocol for the isolation of clones that complement an aft1 mutation in S. cerevisiae that makes cells dependent on iron for growth. After transformation of S. cerevisiae aft1 with a C. albicans library, we have selected clones that grow in conditions of iron deficiency and share an identical plasmid, pIRO1, with a 4500 bp insert containing the URA3 gene and an ORF (IRO1) responsible for the suppression of the iron dependency. IRO1 does not show homology with AFT1 or with other sequences in the databases. Northern analysis demonstrates constitutive expression of IRO1. CAI4, a C. albicans strain isolated as Deltaura3, also has a deletion of the 3' half of IRO1, and displays in YNB medium similar phenotypic characteristics to S. cerevisiae aft1 mutant strains. Therefore, we consider IRO1 as a gene of C. albicans involved in the utilization of iron. However, in extreme conditions of iron deprivation, CAI4 seems to activate alternative mechanisms of iron uptake that allow a better growth than the wild strain SC5314. Analysis of its predicted protein sequence is in agreement with a role of Iro1p as a transcription factor.
García
M G
MG
Departament de Bioquimica i Biologia Molecular, Facultat de Farmacia, Universitat de València, 46100 Burjassot, Spain.
O'Connor
J E
JE
García
L L
LL
Martínez
S I
SI
Herrero
E
E
del Castillo Agudo
L
L
D000595:Amino Acid Sequence; D002176:Candida albicans / Q000166:cytology / Q000235:genetics* / Q000378:metabolism; D005656:Fungal Proteins / Q000235:genetics*; D005800:Genes, Fungal; D008040:Genetic Linkage; D007501:Iron / Q000378:metabolism*; D008969:Molecular Sequence Data; D009964:Orotidine-5'-Phosphate Decarboxylase / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins; D017386:Sequence Homology, Amino Acid; D013489:Suppression, Genetic; D014157:Transcription Factors / Q000235:genetics*
C090297:AFT1 protein, S cerevisiae; D005656:Fungal Proteins; C430663:IRO1 protein, Candida albicans; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D007501:Iron; D009964:Orotidine-5'-Phosphate Decarboxylase