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. 1995 Nov;63(11):4506–4514. doi: 10.1128/iai.63.11.4506-4514.1995

Structure and regulation of the HSP90 gene from the pathogenic fungus Candida albicans.

R K Swoboda 1, G Bertram 1, S Budge 1, G W Gooday 1, N A Gow 1, A J Brown 1
PMCID: PMC173642  PMID: 7591093

Abstract

Candida albicans HSP90 sequences were isolated by screening cDNA and genomic libraries with a probe derived from the Saccharomyces cerevisiae homolog, HSP82, which encodes a member of the heat shock protein 90 family of molecular chaperones. Identical sequences were obtained for the 2,197-bp overlap of the cDNA and gene sequences, which were derived from C. albicans 3153A and ATCC 10261, respectively. The C. albicans HSP90 gene contained no introns, and it showed strong homology (61 to 79% identity) to HSP90 sequences from other fungi, vertebrates, and plants. The C-terminal portion of the predicted Hsp90 amino acid sequence was identical to the 47-kDa protein which is thought to be immunoprotective during C. albicans infections (R. C. Matthews, J. Med. Microbiol. 36:367-370, 1992), confirming that this protein represents the C-terminal portion of the 81-kDa Hsp90 protein. Quantitative Northern (RNA) analyses revealed that C. albicans HSP90 mRNA was heat shock inducible and that its levels changed during batch growth, with its maximum levels being reached during the mid-exponential growth phase. HSP90 mRNA levels increased transiently during the yeast-to-hyphal transition but did not correlate directly with germ tube production per se. These data do not exclude a role for Hsp90 in the dimorphic transition. Southern blotting revealed only one HSP90 locus in the diploid C. albicans genome. Repeated attempts to disrupt both alleles and generate a homozygous C. albicans delta hsp90/delta hsp90 null mutant were unsuccessful. These observations suggest the existence of a single HSP90 locus which is essential for viability in C. albicans.

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Selected References

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  1. Alani E., Cao L., Kleckner N. A method for gene disruption that allows repeated use of URA3 selection in the construction of multiply disrupted yeast strains. Genetics. 1987 Aug;116(4):541–545. doi: 10.1534/genetics.112.541.test. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bonnefoy S., Attal G., Langsley G., Tekaia F., Mercereau-Puijalon O. Molecular characterization of the heat shock protein 90 gene of the human malaria parasite Plasmodium falciparum. Mol Biochem Parasitol. 1994 Sep;67(1):157–170. doi: 10.1016/0166-6851(94)90105-8. [DOI] [PubMed] [Google Scholar]
  3. Borkovich K. A., Farrelly F. W., Finkelstein D. B., Taulien J., Lindquist S. hsp82 is an essential protein that is required in higher concentrations for growth of cells at higher temperatures. Mol Cell Biol. 1989 Sep;9(9):3919–3930. doi: 10.1128/mcb.9.9.3919. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Breathnach R., Chambon P. Organization and expression of eucaryotic split genes coding for proteins. Annu Rev Biochem. 1981;50:349–383. doi: 10.1146/annurev.bi.50.070181.002025. [DOI] [PubMed] [Google Scholar]
  5. Buffo J., Herman M. A., Soll D. R. A characterization of pH-regulated dimorphism in Candida albicans. Mycopathologia. 1984 Mar 15;85(1-2):21–30. doi: 10.1007/BF00436698. [DOI] [PubMed] [Google Scholar]
  6. Cellier M. F., Taimi M., Chateau M. T., Cannat A., Marti J. Thermal stress as an inducer of differentiation of U937 cells. Leuk Res. 1993 Aug;17(8):649–656. doi: 10.1016/0145-2126(93)90069-w. [DOI] [PubMed] [Google Scholar]
  7. Craig E. A., Gambill B. D., Nelson R. J. Heat shock proteins: molecular chaperones of protein biogenesis. Microbiol Rev. 1993 Jun;57(2):402–414. doi: 10.1128/mr.57.2.402-414.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cutforth T., Rubin G. M. Mutations in Hsp83 and cdc37 impair signaling by the sevenless receptor tyrosine kinase in Drosophila. Cell. 1994 Jul 1;77(7):1027–1036. doi: 10.1016/0092-8674(94)90442-1. [DOI] [PubMed] [Google Scholar]
  9. Cutler J. E. Putative virulence factors of Candida albicans. Annu Rev Microbiol. 1991;45:187–218. doi: 10.1146/annurev.mi.45.100191.001155. [DOI] [PubMed] [Google Scholar]
  10. Delbrück S., Ernst J. F. Morphogenesis-independent regulation of actin transcript levels in the pathogenic yeast Candida albicans. Mol Microbiol. 1993 Nov;10(4):859–866. doi: 10.1111/j.1365-2958.1993.tb00956.x. [DOI] [PubMed] [Google Scholar]
  11. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Dhawale S. S., Lane A. C. Compilation of sequence-specific DNA-binding proteins implicated in transcriptional control in fungi. Nucleic Acids Res. 1993 Dec 11;21(24):5537–5546. doi: 10.1093/nar/21.24.5537. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Farrelly F. W., Finkelstein D. B. Complete sequence of the heat shock-inducible HSP90 gene of Saccharomyces cerevisiae. J Biol Chem. 1984 May 10;259(9):5745–5751. [PubMed] [Google Scholar]
  14. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  15. Fonzi W. A., Irwin M. Y. Isogenic strain construction and gene mapping in Candida albicans. Genetics. 1993 Jul;134(3):717–728. doi: 10.1093/genetics/134.3.717. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Gething M. J., Sambrook J. Protein folding in the cell. Nature. 1992 Jan 2;355(6355):33–45. doi: 10.1038/355033a0. [DOI] [PubMed] [Google Scholar]
  17. Hendrick J. P., Hartl F. U. Molecular chaperone functions of heat-shock proteins. Annu Rev Biochem. 1993;62:349–384. doi: 10.1146/annurev.bi.62.070193.002025. [DOI] [PubMed] [Google Scholar]
  18. Jakob U., Buchner J. Assisting spontaneity: the role of Hsp90 and small Hsps as molecular chaperones. Trends Biochem Sci. 1994 May;19(5):205–211. doi: 10.1016/0968-0004(94)90023-x. [DOI] [PubMed] [Google Scholar]
  19. Kaufmann S. H. Heat shock proteins and the immune response. Immunol Today. 1990 Apr;11(4):129–136. doi: 10.1016/0167-5699(90)90050-j. [DOI] [PubMed] [Google Scholar]
  20. Lee K. L., Buckley H. R., Campbell C. C. An amino acid liquid synthetic medium for the development of mycelial and yeast forms of Candida Albicans. Sabouraudia. 1975 Jul;13(2):148–153. doi: 10.1080/00362177585190271. [DOI] [PubMed] [Google Scholar]
  21. Lindquist S., Craig E. A. The heat-shock proteins. Annu Rev Genet. 1988;22:631–677. doi: 10.1146/annurev.ge.22.120188.003215. [DOI] [PubMed] [Google Scholar]
  22. Lindquist S. Regulation of protein synthesis during heat shock. Nature. 1981 Sep 24;293(5830):311–314. doi: 10.1038/293311a0. [DOI] [PubMed] [Google Scholar]
  23. Maresca B., Kobayashi G. S. Dimorphism in Histoplasma capsulatum: a model for the study of cell differentiation in pathogenic fungi. Microbiol Rev. 1989 Jun;53(2):186–209. doi: 10.1128/mr.53.2.186-209.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Matthews R. C., Burnie J. P., Howat D., Rowland T., Walton F. Autoantibody to heat-shock protein 90 can mediate protection against systemic candidosis. Immunology. 1991 Sep;74(1):20–24. [PMC free article] [PubMed] [Google Scholar]
  25. Matthews R. C. Pathogenicity determinants of Candida albicans: potential targets for immunotherapy? Microbiology. 1994 Jul;140(Pt 7):1505–1511. doi: 10.1099/13500872-140-7-1505. [DOI] [PubMed] [Google Scholar]
  26. Matthews R. C. The 14th C. L. Oakley Lecture. Candida albicans HSP 90: link between protective and auto immunity. J Med Microbiol. 1992 Jun;36(6):367–370. doi: 10.1099/00222615-36-6-367. [DOI] [PubMed] [Google Scholar]
  27. Matthews R., Burnie J. Cloning of a DNA sequence encoding a major fragment of the 47 kilodalton stress protein homologue of Candida albicans. FEMS Microbiol Lett. 1989 Jul 1;51(1):25–30. doi: 10.1016/0378-1097(89)90071-2. [DOI] [PubMed] [Google Scholar]
  28. Matthews R., Burnie J., Smith D., Clark I., Midgley J., Conolly M., Gazzard B. Candida and AIDS: evidence for protective antibody. Lancet. 1988 Jul 30;2(8605):263–266. doi: 10.1016/s0140-6736(88)92547-0. [DOI] [PubMed] [Google Scholar]
  29. Moore P. A., Sagliocco F. A., Wood R. M., Brown A. J. Yeast glycolytic mRNAs are differentially regulated. Mol Cell Biol. 1991 Oct;11(10):5330–5337. doi: 10.1128/mcb.11.10.5330. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Picard D., Khursheed B., Garabedian M. J., Fortin M. G., Lindquist S., Yamamoto K. R. Reduced levels of hsp90 compromise steroid receptor action in vivo. Nature. 1990 Nov 8;348(6297):166–168. doi: 10.1038/348166a0. [DOI] [PubMed] [Google Scholar]
  31. Rose D. W., Welch W. J., Kramer G., Hardesty B. Possible involvement of the 90-kDa heat shock protein in the regulation of protein synthesis. J Biol Chem. 1989 Apr 15;264(11):6239–6244. [PubMed] [Google Scholar]
  32. Ryley J. F., Ryley N. G. Candida albicans--do mycelia matter? J Med Vet Mycol. 1990;28(3):225–239. [PubMed] [Google Scholar]
  33. Sanchez E. R., Toft D. O., Schlesinger M. J., Pratt W. B. Evidence that the 90-kDa phosphoprotein associated with the untransformed L-cell glucocorticoid receptor is a murine heat shock protein. J Biol Chem. 1985 Oct 15;260(23):12398–12401. [PubMed] [Google Scholar]
  34. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Santiago T. C., Purvis I. J., Bettany A. J., Brown A. J. The relationship between mRNA stability and length in Saccharomyces cerevisiae. Nucleic Acids Res. 1986 Nov 11;14(21):8347–8360. doi: 10.1093/nar/14.21.8347. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Shapira M., McEwen J. G., Jaffe C. L. Temperature effects on molecular processes which lead to stage differentiation in Leishmania. EMBO J. 1988 Sep;7(9):2895–2901. doi: 10.1002/j.1460-2075.1988.tb03147.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Sherwood J., Gow N. A., Gooday G. W., Gregory D. W., Marshall D. Contact sensing in Candida albicans: a possible aid to epithelial penetration. J Med Vet Mycol. 1992;30(6):461–469. doi: 10.1080/02681219280000621. [DOI] [PubMed] [Google Scholar]
  38. Smith D. J., Cooper M., DeTiani M., Losberger C., Payton M. A. The Candida albicans PMM1 gene encoding phosphomannomutase complements a Saccharomyces cerevisiae sec 53-6 mutation. Curr Genet. 1992 Dec;22(6):501–503. doi: 10.1007/BF00326416. [DOI] [PubMed] [Google Scholar]
  39. Soll D. R. The regulation of cellular differentiation in the dimorphic yeast Candida albicans. Bioessays. 1986 Jul;5(1):5–11. doi: 10.1002/bies.950050103. [DOI] [PubMed] [Google Scholar]
  40. Swoboda R. K., Bertram G., Colthurst D. R., Tuite M. F., Gow N. A., Gooday G. W., Brown A. J. Regulation of the gene encoding translation elongation factor 3 during growth and morphogenesis in Candida albicans. Microbiology. 1994 Oct;140(Pt 10):2611–2616. doi: 10.1099/00221287-140-10-2611. [DOI] [PubMed] [Google Scholar]
  41. Swoboda R. K., Bertram G., Delbrück S., Ernst J. F., Gow N. A., Gooday G. W., Brown A. J. Fluctuations in glycolytic mRNA levels during morphogenesis in Candida albicans reflect underlying changes in growth and are not a response to cellular dimorphism. Mol Microbiol. 1994 Aug;13(4):663–672. doi: 10.1111/j.1365-2958.1994.tb00460.x. [DOI] [PubMed] [Google Scholar]
  42. Swoboda R. K., Bertram G., Hollander H., Greenspan D., Greenspan J. S., Gow N. A., Gooday G. W., Brown A. J. Glycolytic enzymes of Candida albicans are nonubiquitous immunogens during candidiasis. Infect Immun. 1993 Oct;61(10):4263–4271. doi: 10.1128/iai.61.10.4263-4271.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Swoboda R. K., Broadbent I. D., Bertram G., Budge S., Gooday G. W., Gow N. A., Brown A. J. Structure and regulation of a Candida albicans RP10 gene which encodes an immunogenic protein homologous to Saccharomyces cerevisiae ribosomal protein 10. J Bacteriol. 1995 Mar;177(5):1239–1246. doi: 10.1128/jb.177.5.1239-1246.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Swoboda R., Miyasaki S., Greenspan D., Greenspan J. S. Heat-inducible ATP-binding proteins of Candida albicans are recognized by sera of infected patients. J Gen Microbiol. 1993 Dec;139(12):2995–3003. doi: 10.1099/00221287-139-12-2995. [DOI] [PubMed] [Google Scholar]
  45. Van der Ploeg L. H., Giannini S. H., Cantor C. R. Heat shock genes: regulatory role for differentiation in parasitic protozoa. Science. 1985 Jun 21;228(4706):1443–1446. doi: 10.1126/science.4012301. [DOI] [PubMed] [Google Scholar]
  46. Xu Y., Lindquist S. Heat-shock protein hsp90 governs the activity of pp60v-src kinase. Proc Natl Acad Sci U S A. 1993 Aug 1;90(15):7074–7078. doi: 10.1073/pnas.90.15.7074. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Zeuthen M. L., Howard D. H. Thermotolerance and the heat-shock response in Candida albicans. J Gen Microbiol. 1989 Sep;135(9):2509–2518. doi: 10.1099/00221287-135-9-2509. [DOI] [PubMed] [Google Scholar]

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