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. 1994 Feb 1;91(3):922–926. doi: 10.1073/pnas.91.3.922

Candida albicans estrogen-binding protein gene encodes an oxidoreductase that is inhibited by estradiol.

N D Madani 1, P J Malloy 1, P Rodriguez-Pombo 1, A V Krishnan 1, D Feldman 1
PMCID: PMC521425  PMID: 8302868

Abstract

Candida albicans, the most common fungal pathogen of humans, possesses an estrogen-binding protein (EBP) that binds mammalian estrogens with high affinity. We report here the cloning and complete nucleotide sequence of a gene encoding a C. albicans EBP. Amino acid sequences obtained from cyanogen bromide fragments of purified EBP were used to design oligonucleotide primers for PCR. An 800-bp product was amplified and used to screen a C. albicans genomic library. A clone was isolated containing an insert with an open reading frame of 1221 nt capable of encoding a protein with 407 amino acids and having a calculated molecular mass of 46,073 Da, the estimated size of EBP. The cloned gene, expressed in Escherichia coli as a lacZ fusion protein, demonstrated high-affinity binding for estradiol and a competition profile comparable to C. albicans wild-type EBP. Northern blots of C. albicans RNA revealed a single transcript of approximately 1600 nt, whereas Southern blots identified three hybridizing fragments. Computer searches of data bases showed that EBP shares a 46% amino acid identity with the old yellow enzyme, an oxidoreductase from Saccharomyces cerevisiae, but was unrelated to the human estrogen receptor as previously speculated. In addition, a 51-amino acid region of EBP is highly conserved among a group of flavoproteins including old yellow enzyme. Expressed EBP was shown to exhibit oxidoreductase activity that could be inhibited by 17 beta-estradiol in vitro. In conclusion, the EBP from C. albicans has no evident homology to the mammalian steroid receptor superfamily but appears to be a member of a recently identified family of flavoproteins.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Abramovitz A. S., Massey V. Purification of intact old yellow enzyme using an affinity matrix for the sole chromatographic step. J Biol Chem. 1976 Sep 10;251(17):5321–5326. [PubMed] [Google Scholar]
  2. Boyd G., Mathews F. S., Packman L. C., Scrutton N. S. Trimethylamine dehydrogenase of bacterium W3A1. Molecular cloning, sequence determination and over-expression of the gene. FEBS Lett. 1992 Aug 24;308(3):271–276. doi: 10.1016/0014-5793(92)81291-s. [DOI] [PubMed] [Google Scholar]
  3. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  4. Brenner C., Fuller R. S. Structural and enzymatic characterization of a purified prohormone-processing enzyme: secreted, soluble Kex2 protease. Proc Natl Acad Sci U S A. 1992 Feb 1;89(3):922–926. doi: 10.1073/pnas.89.3.922. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Burshell A., Stathis P. A., Do Y., Miller S. C., Feldman D. Characterization of an estrogen-binding protein in the yeast Saccharomyces cerevisiae. J Biol Chem. 1984 Mar 25;259(6):3450–3456. [PubMed] [Google Scholar]
  6. Clemons K. V., Schär G., Stover E. P., Feldman D., Stevens D. A. Dermatophyte-hormone relationships: characterization of progesterone-binding specificity and growth inhibition in the genera Trichophyton and Microsporum. J Clin Microbiol. 1988 Oct;26(10):2110–2115. doi: 10.1128/jcm.26.10.2110-2115.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Clemons K. V., Stover E. P., Schär G., Stathis P. A., Chan K., Tökès L., Stevens D. A., Feldman D. Steroid metabolism as a mechanism of escape from progesterone-mediated growth inhibition in Trichophyton mentagrophytes. J Biol Chem. 1989 Jul 5;264(19):11186–11192. [PubMed] [Google Scholar]
  8. Feldman D., Do Y., Burshell A., Stathis P., Loose D. S. An estrogen-binding protein and endogenous ligand in Saccharomyces cerevisiae: possible hormone receptor system. Science. 1982 Oct 15;218(4569):297–298. doi: 10.1126/science.6289434. [DOI] [PubMed] [Google Scholar]
  9. Kinsman O. S., Pitblado K., Coulson C. J. Effect of mammalian steroid hormones and luteinizing hormone on the germination of Candida albicans and implications for vaginal candidosis. Mycoses. 1988 Dec;31(12):617–626. doi: 10.1111/j.1439-0507.1988.tb04416.x. [DOI] [PubMed] [Google Scholar]
  10. Loose D. S., Feldman D. Characterization of a unique corticosterone-binding protein in Candida albicans. J Biol Chem. 1982 May 10;257(9):4925–4930. [PubMed] [Google Scholar]
  11. Loose D. S., Schurman D. J., Feldman D. A corticosteroid binding protein and endogenous ligand in C. albicans indicating a possible steroid-receptor system. Nature. 1981 Oct 8;293(5832):477–479. doi: 10.1038/293477a0. [DOI] [PubMed] [Google Scholar]
  12. Loose D. S., Stover E. P., Restrepo A., Stevens D. A., Feldman D. Estradiol binds to a receptor-like cytosol binding protein and initiates a biological response in Paracoccidioides brasiliensis. Proc Natl Acad Sci U S A. 1983 Dec;80(24):7659–7663. doi: 10.1073/pnas.80.24.7659. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Maeda K., Truscott K., Liu X. L., Scopes R. K. A thermostable NADH oxidase from anaerobic extreme thermophiles. Biochem J. 1992 Jun 1;284(Pt 2):551–555. doi: 10.1042/bj2840551. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Mallonee D. H., White W. B., Hylemon P. B. Cloning and sequencing of a bile acid-inducible operon from Eubacterium sp. strain VPI 12708. J Bacteriol. 1990 Dec;172(12):7011–7019. doi: 10.1128/jb.172.12.7011-7019.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Malloy P. J., Zhao X., Madani N. D., Feldman D. Cloning and expression of the gene from Candida albicans that encodes a high-affinity corticosteroid-binding protein. Proc Natl Acad Sci U S A. 1993 Mar 1;90(5):1902–1906. doi: 10.1073/pnas.90.5.1902. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Morohashi K., Fujii-Kuriyama Y., Okada Y., Sogawa K., Hirose T., Inayama S., Omura T. Molecular cloning and nucleotide sequence of cDNA for mRNA of mitochondrial cytochrome P-450(SCC) of bovine adrenal cortex. Proc Natl Acad Sci U S A. 1984 Aug;81(15):4647–4651. doi: 10.1073/pnas.81.15.4647. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Murphy J. W. Mechanisms of natural resistance to human pathogenic fungi. Annu Rev Microbiol. 1991;45:509–538. doi: 10.1146/annurev.mi.45.100191.002453. [DOI] [PubMed] [Google Scholar]
  18. Nikodem V., Fresco J. R. Protein fingerprinting by SDS-gel electrophoresis after partial fragmentation with CNBr. Anal Biochem. 1979 Sep 1;97(2):382–386. doi: 10.1016/0003-2697(79)90089-7. [DOI] [PubMed] [Google Scholar]
  19. Restrepo A., Salazar M. E., Cano L. E., Stover E. P., Feldman D., Stevens D. A. Estrogens inhibit mycelium-to-yeast transformation in the fungus Paracoccidioides brasiliensis: implications for resistance of females to paracoccidioidomycosis. Infect Immun. 1984 Nov;46(2):346–353. doi: 10.1128/iai.46.2.346-353.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Saito K., Thiele D. J., Davio M., Lockridge O., Massey V. The cloning and expression of a gene encoding Old Yellow Enzyme from Saccharomyces carlsbergensis. J Biol Chem. 1991 Nov 5;266(31):20720–20724. [PubMed] [Google Scholar]
  21. Schär G., Stover E. P., Clemons K. V., Feldman D., Stevens D. A. Progesterone binding and inhibition of growth in Trichophyton mentagrophytes. Infect Immun. 1986 Jun;52(3):763–767. doi: 10.1128/iai.52.3.763-767.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Skowronski R., Feldman D. Characterization of an estrogen-binding protein in the yeast Candida albicans. Endocrinology. 1989 Apr;124(4):1965–1972. doi: 10.1210/endo-124-4-1965. [DOI] [PubMed] [Google Scholar]
  23. Stott K., Saito K., Thiele D. J., Massey V. Old Yellow Enzyme. The discovery of multiple isozymes and a family of related proteins. J Biol Chem. 1993 Mar 25;268(9):6097–6106. [PubMed] [Google Scholar]
  24. Tanaka S., Hasegawa S., Hishinuma F., Kurata S. Estrogen can regulate the cell cycle in the early G1 phase of yeast by increasing the amount of adenylate cyclase mRNA. Cell. 1989 May 19;57(4):675–681. doi: 10.1016/0092-8674(89)90136-0. [DOI] [PubMed] [Google Scholar]
  25. Yielding K. L., Tomkins G. M. STRUCTURAL ALTERATIONS IN CRYSTALLINE GLUTAMIC DEHYDROGENASE INDUCED BY STEROID HORMONES. Proc Natl Acad Sci U S A. 1960 Nov;46(11):1483–1488. doi: 10.1073/pnas.46.11.1483. [DOI] [PMC free article] [PubMed] [Google Scholar]

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