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

Infect Immun. 1995 November; 63(11): 4506–4514.

PMCID: PMC173642

R K Swoboda, G Bertram, S Budge, G W Gooday, N A Gow, and A J Brown

Department of Molecular and Cell Biology, Marischal College, University of Aberdeen, United Kingdom.

This article has been corrected. See Infect Immun. 1996 February; 64(2): 680.

This article has been cited by other articles in PMC.

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.

Full Text

The Full Text of this article is available as a PDF (1.0M).

Selected References

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

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. [PubMed]
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. [PubMed]
Borkovich, KA; Farrelly, FW; Finkelstein, DB; 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. [PubMed]
Breathnach, R; Chambon, P. Organization and expression of eucaryotic split genes coding for proteins. Annu Rev Biochem. 1981;50:349–383. [PubMed]
Buffo, J; Herman, MA; Soll, DR. A characterization of pH-regulated dimorphism in Candida albicans. Mycopathologia. 1984 Mar 15;85(1-2):21–30. [PubMed]
Cellier, MF; Taimi, M; Chateau, MT; Cannat, A; Marti, J. Thermal stress as an inducer of differentiation of U937 cells. Leuk Res. 1993 Aug;17(8):649–656. [PubMed]
Craig, EA; Gambill, BD; Nelson, RJ. Heat shock proteins: molecular chaperones of protein biogenesis. Microbiol Rev. 1993 Jun;57(2):402–414. [PubMed]
Cutforth, T; Rubin, GM. Mutations in Hsp83 and cdc37 impair signaling by the sevenless receptor tyrosine kinase in Drosophila. Cell. 1994 Jul 1;77(7):1027–1036. [PubMed]
Cutler, JE. Putative virulence factors of Candida albicans. Annu Rev Microbiol. 1991;45:187–218. [PubMed]
Delbrück, S; Ernst, JF. Morphogenesis-independent regulation of actin transcript levels in the pathogenic yeast Candida albicans. Mol Microbiol. 1993 Nov;10(4):859–866. [PubMed]
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. [PubMed]
Dhawale, SS; Lane, AC. Compilation of sequence-specific DNA-binding proteins implicated in transcriptional control in fungi. Nucleic Acids Res. 1993 Dec 11;21(24):5537–5546. [PubMed]
Farrelly, FW; Finkelstein, DB. Complete sequence of the heat shock-inducible HSP90 gene of Saccharomyces cerevisiae. J Biol Chem. 1984 May 10;259(9):5745–5751. [PubMed]
Feinberg, AP; Vogelstein, B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. [PubMed]
Fonzi, WA; Irwin, MY. Isogenic strain construction and gene mapping in Candida albicans. Genetics. 1993 Jul;134(3):717–728. [PubMed]
Gething, MJ; Sambrook, J. Protein folding in the cell. Nature. 1992 Jan 2;355(6355):33–45. [PubMed]
Hendrick, JP; Hartl, FU. Molecular chaperone functions of heat-shock proteins. Annu Rev Biochem. 1993;62:349–384. [PubMed]
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. [PubMed]
Kaufmann, SH. Heat shock proteins and the immune response. Immunol Today. 1990 Apr;11(4):129–136. [PubMed]
Lee, KL; Buckley, HR; Campbell, CC. An amino acid liquid synthetic medium for the development of mycelial and yeast forms of Candida Albicans. Sabouraudia. 1975 Jul;13(2):148–153. [PubMed]
Lindquist, S. Regulation of protein synthesis during heat shock. Nature. 1981 Sep 24;293(5830):311–314. [PubMed]
Lindquist, S; Craig, EA. The heat-shock proteins. Annu Rev Genet. 1988;22:631–677. [PubMed]
Maresca, B; Kobayashi, GS. Dimorphism in Histoplasma capsulatum: a model for the study of cell differentiation in pathogenic fungi. Microbiol Rev. 1989 Jun;53(2):186–209. [PubMed]
Matthews, RC. 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. [PubMed]
Matthews, RC. Pathogenicity determinants of Candida albicans: potential targets for immunotherapy? Microbiology. 1994 Jul;140 (:1505–1511. [PubMed]
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. [PubMed]
Matthews, RC; Burnie, JP; 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. [PubMed]
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. [PubMed]
Moore, PA; Sagliocco, FA; Wood, RM; Brown, AJ. Yeast glycolytic mRNAs are differentially regulated. Mol Cell Biol. 1991 Oct;11(10):5330–5337. [PubMed]
Picard, D; Khursheed, B; Garabedian, MJ; Fortin, MG; Lindquist, S; Yamamoto, KR. Reduced levels of hsp90 compromise steroid receptor action in vivo. Nature. 1990 Nov 8;348(6297):166–168. [PubMed]
Rose, DW; Welch, WJ; 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]
Ryley, JF; Ryley, NG. Candida albicans--do mycelia matter? J Med Vet Mycol. 1990;28(3):225–239. [PubMed]
Sanchez, ER; Toft, DO; Schlesinger, MJ; Pratt, WB. 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]
Sanger, F; Nicklen, S; Coulson, AR. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. [PubMed]
Santiago, TC; Purvis, IJ; Bettany, AJ; Brown, AJ. The relationship between mRNA stability and length in Saccharomyces cerevisiae. Nucleic Acids Res. 1986 Nov 11;14(21):8347–8360. [PubMed]
Shapira, M; McEwen, JG; Jaffe, CL. Temperature effects on molecular processes which lead to stage differentiation in Leishmania. EMBO J. 1988 Sep;7(9):2895–2901. [PubMed]
Sherwood, J; Gow, NA; Gooday, GW; Gregory, DW; Marshall, D. Contact sensing in Candida albicans: a possible aid to epithelial penetration. J Med Vet Mycol. 1992;30(6):461–469. [PubMed]
Smith, DJ; Cooper, M; DeTiani, M; Losberger, C; Payton, MA. The Candida albicans PMM1 gene encoding phosphomannomutase complements a Saccharomyces cerevisiae sec 53-6 mutation. Curr Genet. 1992 Dec;22(6):501–503. [PubMed]
Soll, DR. The regulation of cellular differentiation in the dimorphic yeast Candida albicans. Bioessays. 1986 Jul;5(1):5–11. [PubMed]
Swoboda, RK; Bertram, G; Colthurst, DR; Tuite, MF; Gow, NA; Gooday, GW; Brown, AJ. Regulation of the gene encoding translation elongation factor 3 during growth and morphogenesis in Candida albicans. Microbiology. 1994 Oct;140 (:2611–2616. [PubMed]
Swoboda, RK; Bertram, G; Delbrück, S; Ernst, JF; Gow, NA; Gooday, GW; Brown, AJ. 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. [PubMed]
Swoboda, RK; Bertram, G; Hollander, H; Greenspan, D; Greenspan, JS; Gow, NA; Gooday, GW; Brown, AJ. Glycolytic enzymes of Candida albicans are nonubiquitous immunogens during candidiasis. Infect Immun. 1993 Oct;61(10):4263–4271. [PubMed]
Swoboda, RK; Broadbent, ID; Bertram, G; Budge, S; Gooday, GW; Gow, NA; Brown, AJ. 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. [PubMed]
Swoboda, R; Miyasaki, S; Greenspan, D; Greenspan, JS. Heat-inducible ATP-binding proteins of Candida albicans are recognized by sera of infected patients. J Gen Microbiol. 1993 Dec;139(12):2995–3003. [PubMed]
Van der Ploeg, LH; Giannini, SH; Cantor, CR. Heat shock genes: regulatory role for differentiation in parasitic protozoa. Science. 1985 Jun 21;228(4706):1443–1446. [PubMed]
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. [PubMed]
Zeuthen, ML; Howard, DH. Thermotolerance and the heat-shock response in Candida albicans. J Gen Microbiol. 1989 Sep;135(9):2509–2518. [PubMed]