1887

Abstract

The temporal sequence of cytosolic protein expression during phase transition of was examined. Electrophoretic analysis of cytosol proteins by one-dimensional SDS-PAGE revealed numerous differences between the mycelial and yeast forms as well as alterations induced by 17-oestradiol. Using either protein staining or fluorography of [S]methionine-labelled proteins 30 phase-specific bands were detected, 12 mycelial-associated bands (range 30 to 140 kDa) and 18 yeast-associated bands (range 22 to 127 kDa). In cells undergoing mycelial to yeast transition after a shift from 25 °C to 37 °C, the protein patterns showed a temporal progression toward the yeast profile with the accumulation of yeast bands prior to observable morphogenesis. Five novel protein bands (range 23 to 50 kDa) were detected by silver staining during transition. Treatment of temperature-shifted mycelial cultures with 26 × 10 -oestradiol altered observed profiles; 4 of 12 mycelial-associated bands were maintained whereas the appearance of the 5 novel transition bands and 9 of 18 yeast-associated bands was blocked or delayed. Analysis of [S]methionine-labelled proteins revealed that oestradiol induced label uptake by mycelial cells, blocked the synthesis of a 92 kDa yeast-specific band 72 h into transition, and diminished label incorporation 120 h into transition. In conjunction with these steroid-induced alterations of protein expression, little or no morphological transformation occurred. These results support our hypothesis that, analogous to mammalian steroid receptor action, the functional responses of to oestradiol are related to regulation of protein expression, presumably mediated via a specific binding proteinligand complex.

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1989-06-01
2021-07-28
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References

  1. Bonner W. M., Laskey R. A. 1974; A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. European Journal of Biochemistry 46:83–88
    [Google Scholar]
  2. Bradford M. M. 1976; A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Annals of Biochemistry 72:248–254
    [Google Scholar]
  3. Brunt S. A., Silver J. C. 1986a; Steroid hormone-induced changes in secreted proteins in the filamentous fungus Achlya. Experimental Cell Research 16322–34
    [Google Scholar]
  4. Brunt S. A., Silver J. C. 1986b; Cellular localization of steroid hormone-regulated proteins during sexual development in Achlya. Experimental Cell Research 165:306–319
    [Google Scholar]
  5. Brunt S. A., Silver J. C. 1987; Steroid hormone-regulated basic proteins in Achlya ambisexualis. Experimental Mycology 11:65–69
    [Google Scholar]
  6. Carbonell L., Rodriguez J. 1965; Transformation of mycelial and yeast forms of Paracoccidioides brasiliensis in cultures and in experimental inoculations. Journal of Bacteriology 90:504–510
    [Google Scholar]
  7. Dabrowa N., Howard D. H. 1984; Heat shock and heat stroke proteins observed during germination of the blastoconidia of Candida albicans. Infection and Immunity 44:537–539
    [Google Scholar]
  8. Kane J. 1984; Conversion of Blastomyces dermatitidis to the yeast form at 37 °C and 26 °C. Journal of Clinical Microbiology 20:594–596
    [Google Scholar]
  9. Kanetsuna F., Carbonell L. M., Azuma I., Yamamura Y. 1972; Biochemical studies on the thermal dimorphism of Paracoccidioides brasiliensis. Journal of Bacteriology 110:208–218
    [Google Scholar]
  10. Kobayashi G. S., Medoff G., Maresca B., Sacco M., Kumar B. V. 1985; Studies on phase transitions in the dimorphic pathogen Histoplasma capsulatum. In Fungal Dimorphism with Emphasis on Fungi Pathogenic for Humans pp. 69–91 Szaniszlo P. J., Harris J. L. Edited by New York: Plenum Press;
    [Google Scholar]
  11. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature; London: 227680–685
    [Google Scholar]
  12. Lambowitz A. M., Kobayashi G. S., Painter A., Medoff G. 1983; Possible relationship of morphogenesis in pathogenic fungus, Histoplasma capsulatum, to heat shock response. Nature; London: 303806–808
    [Google Scholar]
  13. Loose D. S., Stover E. P., Restrepo A., Stevens D. A., Feldman D. 1983; Estradiol binds to a receptor-like cytosol binding protein and initiates a biological response in Paracoccidioides brasiliensis. Proceedings of the National Academy of Sciences of the United States of America 80:7659–7663
    [Google Scholar]
  14. Maresca B., Lambowitz A. M., Kumar V. B., Grant G. A., Kobayashi G. S., Medoff G. 1981; Role of cysteine in regulating morphogenesis and mitochondrial activity in the dimorphic fungus Histoplasma capsulatum. Proceedings of the National Academy of Sciences of the United States of America 78:4596–4600
    [Google Scholar]
  15. Medoff G., Kobayashi G. S., Painter A., Travis S. 1987a; Morphogenesis and pathogenicity of Histoplasma capsulatum. Infection and Immunity 55:1355–1358
    [Google Scholar]
  16. Medoff G., Painter A., Kobayashi G. S. 1987b; Mycelial-to-yeast-phase transitions of the dimorphic fungi Blastomyces dermatitidis and Paracoccidioides brasiliensis. Journal of Bacteriology 169:4055–4060
    [Google Scholar]
  17. Miyakawa T., Kadota T., Okubo Y., Hatano T., Tsuchiya E., Fukui S. 1984; Mating pheromone-induced alteration of cell surface proteins in the heterobasidiomycetous yeast Tremella mesenterica. Journal of Bacteriology 158:814–819
    [Google Scholar]
  18. Neidhardt F. C., Van Bogelen R. A., Vaughn V. 1984; The genetics and regulation of heat-shock proteins. Annual Review of Genetics 18:295–329
    [Google Scholar]
  19. Paris S., Duran S. 1985; Cyclic adenosine 3′,5′-monophosphate (cAMP) and dimorphism in the pathogenic fungus Paracoccidioides brasiliensis. Mycopathologia 92:115–120
    [Google Scholar]
  20. Paris S., Duran-Gonzalez S., Mariat F. 1985; Nutritional studies on Paracoccidioides brasiliensis:the role of organic sulfur in dimorphism. Sabouraudia 23:85–92
    [Google Scholar]
  21. Patino M. M., Burgas L. C., Restrepo A. 1984; Effect of temperature on the mycelium to yeast transformation of Paracoccidioides brasiliensis. Sabouraudia 22:509–511
    [Google Scholar]
  22. Restrepo M. A. 1985; The ecology of Paracoccidioides brasiliensis: a puzzle still unsolved. Sabouraudia 23:323–334
    [Google Scholar]
  23. Restrepo A., Jimenez B. E. 1980; Growth of Paracoccidioides brasiliensis yeast phase in a chemically defined culture medium. Journal of Clinical Microbiology 12:279–281
    [Google Scholar]
  24. Restrepo A., Salazar M. E., Cano L. E., Stover E. P., Feldman D., Stevens D. A. 1984; Estrogens inhibit mycelium-to-yeast transformation in the fungus Paracoccidioides brasiliensis: implications for resistance of females to paracoccidioidomycosis. Infection and Immunity 46:346–353
    [Google Scholar]
  25. Riehl R. M., Toft D. O. 1984; Analysis of the steroid receptor of Achlya ambisexualis. Journal of Biological Chemistry 259:15324–15330
    [Google Scholar]
  26. Riehl R. M., Toft D. O., Meyer M. D., Carlson G. L., Mcmorris T. C. 1984; Detection of a pheromone-binding protein in the aquatic fungus Achlya ambisexualis. Experimental Cell Research 153:544–549
    [Google Scholar]
  27. Salazar M. E., Restrepo A. 1984; Morphogenesis of the mycelium to yeast transformation in Paracoccidioides brasiliensis. Sabouraudia 22:7–11
    [Google Scholar]
  28. San-Blas F., San-Blas G. 1985 Paracoccidioides brasiliensis. In Fungal Dimorphism with Emphasis on Fungi Pathogenic for Humans pp. 93–120 Szaniszlo P. J., Harris J. L. Edited by New York: Plenum Press;
    [Google Scholar]
  29. San-Blas G., Vernet D. 1977; Induction of the synthesis of cell wall α1,3-glucan in the yeastlike form of Paracoccidioides brasiliensis strain IVIC Pb9 by fetal calf serum. Infection and Immunity 15:897–902
    [Google Scholar]
  30. Stover E. P., Schar G., Clemons K. V., Stevens D. A., Feldman D. 1986; Estradiol-binding proteins from mycelial and yeast-form cultures of Paracoccidioides brasiliensis. Infection and Immunity 51:199–203
    [Google Scholar]
  31. Yamamoto K. R. 1985; Steroid receptor regulated transcription of specific genes and gene networks. Annual Review of Genetics 19:209–252
    [Google Scholar]
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