A high-affinity iron permease essential for Candida albicans virulence

Science

Volumn 288, Issue 5468, 2000, Pages 1062-1064

  Ramanan, Narendrakumar ^a   Wang, Yue ^a  

^a INSTITUTE OF MOLECULAR AND CELL BIOLOGY   (Singapore)

Author keywords

[No Author keywords available]

Indexed keywords

FUNGAL ENZYME; IRON; PERMEASE;

ANIMAL CELL; ANIMAL MODEL; ARTICLE; CANDIDA ALBICANS; CANDIDIASIS; FUNGUS GROWTH; GENE EXPRESSION; GENE EXPRESSION REGULATION; GENE INDUCTION; GENE ISOLATION; GENE REPRESSION; IRON TRANSPORT; MOUSE; NONHUMAN; PRIORITY JOURNAL; VIRULENCE;

ANIMALS; BIOLOGICAL TRANSPORT; CANDIDA ALBICANS; CANDIDIASIS; CARRIER PROTEINS; CELL MEMBRANE; CULTURE MEDIA; FERRIC COMPOUNDS; GENE DELETION; GENE EXPRESSION REGULATION, FUNGAL; GENES, FUNGAL; IRON; KIDNEY; MEMBRANE TRANSPORT PROTEINS; MICE; RECOMBINANT FUSION PROTEINS; SACCHAROMYCES CEREVISIAE; SACCHAROMYCES CEREVISIAE PROTEINS; TRANSFORMATION, GENETIC; VIRULENCE;

EID: 0034640466     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.288.5468.1062     Document Type: Article

References (33)

1. M. J. Richards, J. R. Edwards, D. H. Culver, R. P. Gaynes, Crit. Care Med. 27, 887 (1999).
2. H. J. Lo et al., Cell 90, 939 (1997).
3. D. Sanglard, B. Hube, M. Monod, F. C. Odds, N. A. Gow, Infect. Immun. 65, 3539 (1997).
4. C. A. Gale et al., Science 279, 1355 (1998).
5. C. C. Askwith and J. Kaplan, Trends Biochem. Sci. 23, 135 (1998).
6. S. M. Vidal et al., Cell 73, 469 (1993).
7. D. M. De Silva, C. C. Askwith, J. Kaplan, Physiol. Rev. 76, 31 (1996).
8. H. Gunshin et al., Nature 388, 482 (1997).
9. M. K. Stevens et al., Infect. Immun. 64, 1724 (1996).
10. C. N. Cornelissen et al., Mol. Microbiol. 27, 611(1998).
11. A. B. Schryvers and I. Stojiljkovic, Mol. Microbiol. 32, 1117 (1999).
12. - strains in SD, 50 μg/ml uridine was added. From C. albicans strain (ATCC 10259), two cultures grown in LlM2 and LlM1000 were used to construct a subtracted cDNA library to enrich genes differentially expressed in LlM2 by using the polymerase chain reaction (PCR) Select Subtracted cDNA Library kit (Clontech). Sequencing the library clones identified two overlapping cDNA of 160 and 210 base pairs (bp). The cDNA was used to probe a C. albicans lambda genomic library Two DNA fragments of ∼5.7 and 5.4 kbp, encoding CaFTR1 and CaFTR2, respectively, were isolated and sequenced using a DNA sequencer (Applied Biosystems, model 377).
13. R. Stearman, D. S. Yuan, Y. Yamaguchi-Iwai, R. D. Klausner, A. Dancis, Science 271, 1552 (1996).
14. Web supplementary material is available to Science Online subscribers at www.sciencemag.org/feature/ data/1047970.shl.
15. 32P-labeled as probes. Blots were washed in 0.2× standard saline citrate with 0.1% SDS for 1 hour at 65°C.
16. S. cerevisiae haploid strain CRY2α (can1-100, ade2-1, his3-11, 15, leu2-3, 112, trp1-1, ura3) was used to construct the ftr1 strain (14).
17. D. Eide et al., J. Biol. Chem. 267, 20774 (1992).
18. 3 (2 μM) was used in a 10-minute assay at 30°C. Radioactivity was counted by a LKB Compugamma counter (model 1282), and the amount bound to the cells kept on ice was subtracted as background.
19. C. Askwith and J. Kaplan, J. Biol. Chem. 272, 401 (1997).
20. N. Ramanan and Y. Wang, data not shown.
21. E. Georgatsou and D. Alexandraki, Mol. Cell. Biol. 14, 3065 (1994).
22. R. Eck, S. Hundt, A. Hartl, E. Roemer, W. Kunkel, Microbiology 145, 2415 (1999).
23. - Caftr1 strain and transformants selected on SD200-uridine plates.
24. J. R. Kohler and G. R. Fink, Proc. Natl. Acad. Sci. U.S.A. 93, 13223 (1996).
25. To construct a plasmid expressing CaFTR2, primers 5′-tcatacgtcgattaataa-3′ and 5′-gcggatcctgttaatgtttatttttt-3′ amplified CaFTR1 promoter [nucleotides (nt) -1292 to -1, the first base of the start codon ATG is nt 1] from pABFTR1 with a Bam HI site added on the 3′ end. The PCR fragment was cut with Cla I (nt -1287) and Bam HI and inserted between the corresponding sites in pABSK1 to yield pP1. The CaFTR2 coding region and 400-bp 3′ flanking sequence (nt 1 to 1479) was amplified by using a high-fidelity PCR kit (Roche) from the genomic clone with Bam HI and Xba I sites on the 5′ and 3′ ends, respectively. This PCR fragment and pP1 were cut with Bam HI and Xba I and ligated to yield pP1O2.
26. W. A. Fonzi, J. Bacteriol. 181, 7070 (1999).
27. - Caftr1 cells and clones selected on SD200-uridine plates. The transformants were grown in either SD50 or SD1000 to exponential phase for fluorescence microscopy (Leica).
28. N. Ramanan and Y. Wang, data not shown.
29. D. H. Howard, Clin. Microbiol. Rev. 12, 394 (1999).
30. D. Eide and L Guarente, J. Gen. Microbiol. 138, 347 (1992).
31. D. J. D. Nicholas, Methods Enzymol. 3, 1035 (1965).
32. B. P. Cormack et al., Microbiology 143, 303 (1997).
33. This work was supported by the Institute of Molecular and Cell Biology. We thank J. Kaplan for advice on setting up the iron-uptake assay, W. J. Hong, U. Surana, C. J. Pallen, A. E. Ting and P. Singh for critically reading the manuscript, U. Surana's and M. J. Cai's laboratories for help with yeast experiments, R. M. L. Choong for help with animal experiments, M. Balasubramanian for the use of fluorescence microscope, and W. Fonzi and D. Brown for providing Candida strains.