1887

Abstract

is a pathogenic fungus that produces melanin when incubated in the presence of certain phenolic substrates such as -3,4-dihydroxyphenylalanine (-dopa). Melanin is an enigmatic polymer that is deposited in the cell wall and contributes to virulence. Substantial progress has been made in understanding the synthesis of melanin and the mechanisms by which it contributes to virulence, but relatively little is known about how melanin is rearranged during growth and budding. In this study we used transmission and scanning electron microscopy and immunofluorescence of melanized cells and melanin ‘ghosts' to study the process of melanization during replication. Budding in melanized results in focal disruption of cell-wall melanin at the bud site. In the presence of -dopa, bud-related melanin defects are repaired and daughter cells are melanized. However, in the absence of substrate, mother cells cannot repair their melanin defects and daughter cells are non-melanized. Hence, melanin in the parent cell is not carried to the daughter cells, but rather is synthesized in buds. These results imply that melanin remodelling occurs during cell growth in a process that involves degradation and synthesis at sites of budding.

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2003-07-01
2019-10-19
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References

  1. Alspaugh, J. A., Perfect, J. R. & Heitman, J. ( 1997; ). Cryptococcus neoformans mating and virulence are regulated by the G-protein alpha subunit GPA1 and cAMP. Genes Dev 11, 3206–3217.[CrossRef]
    [Google Scholar]
  2. Alspaugh, J. A., Perfect, J. R. & Heitman, J. ( 1998; ). Signal transduction pathways regulating differentiation and pathogenicity of Cryptococcus neoformans. Fungal Genet Biol 25, 1–14.[CrossRef]
    [Google Scholar]
  3. Bell, A. A. & Wheeler, M. H. ( 1986; ). Biosynthesis and functions of fungal melanins. Annu Rev Phytopathol 24, 411–451.[CrossRef]
    [Google Scholar]
  4. Brush, L. & Money, N. P. ( 1999; ). Invasive hyphal growth in Wangiella dermatitidis is induced by stab inoculation and shows dependence upon melanin biosynthesis. Fungal Genet Biol 28, 190–200.[CrossRef]
    [Google Scholar]
  5. Butler, M. J. & Day, A. W. ( 1998; ). Ligninases are melaninases. Int J Plant Sci 159, 989–995.[CrossRef]
    [Google Scholar]
  6. Cassone, A., Simonetti, N. & Strippoli, V. ( 1974; ). Wall structure and bud formation on Cryptococcus neoformans. Arch Microbiol 95, 205–212.[CrossRef]
    [Google Scholar]
  7. Chen, L. C., Blank, E. S. & Casadevall, A. ( 1996; ). Extracellular proteinase activity of Cryptococcus neoformans. Clin Diagn Lab Immunol 3, 570–574.
    [Google Scholar]
  8. Chen, L. C., Pirofski, L. A. & Casadevall, A. ( 1997; ). Extracellular proteins of Cryptococcus neoformans and host antibody response. Infect Immun 65, 2599–2605.
    [Google Scholar]
  9. Cid, V. J., Jiménez, J., Molina, M., Sánchez, M., Nombela, C. & Thorner, J. W. ( 2002; ). Orchestrating the cell cycle in yeast: sequential localization of key mitotic regulators at the spindle pole and the bud neck. Microbiology 148, 2647–2659.
    [Google Scholar]
  10. Dixon, D. M., Polak, A. & Conner, G. W. ( 1989; ). Mel mutants of Wangiella dermatitidis in mice: evaluation of multiple mouse and fungal strains. J Med Vet Mycol 27, 335–341.[CrossRef]
    [Google Scholar]
  11. Dixon, D. M., Szaniszlo, P. J. & Polak, A. ( 1991; ). Dihydroxynaphthalene (DHN) melanin and its relationship with virulence in the early stages of phaeohyphomycosis. In The Fungal Spore and Disease Initiation in Plants and Animals, pp. 297–318. Edited by G. Cole & H. Hoch. New York: Plenum.
  12. Doering, T. L., Nosanchuk, J. D., Roberts, W. K. & Casadevall, A. ( 1999; ). Melanin as a potential cryptococcal defence against microbicidal proteins. Med Mycol 37, 175–181.[CrossRef]
    [Google Scholar]
  13. Drees, B. L., Sundin, B., Brazeau, E. & 19 other authors ( 2001; ). A protein interaction map for cell polarity development. J Cell Biol 154, 549–571.[CrossRef]
    [Google Scholar]
  14. D'Souza, C. A., Alspaugh, J. A., Yue, C., Harashima, T., Cox, G. M., Perfect, J. R. & Heitman, J. ( 2001; ). Cyclic AMP-dependent protein kinase controls virulence of the fungal pathogen Cryptococcus neoformans. Mol Cell Biol 21, 3179–3191.[CrossRef]
    [Google Scholar]
  15. Eggert, C., Temp, U. & Eriksson, K. E. ( 1996a; ). The ligninolytic system of the white rot fungus Pycnoporus cinnabarinus: purification and characterization of the laccase. Appl Environ Microbiol 62, 1151–1158.
    [Google Scholar]
  16. Eggert, C., Temp, U., Dean, J. F. & Eriksson, K. E. ( 1996b; ). A fungal metabolite mediates degradation of non-phenolic lignin structures and synthetic lignin by laccase. FEBS Lett 391, 144–148.[CrossRef]
    [Google Scholar]
  17. Eggert, C., Temp, U. & Eriksson, K. E. ( 1997; ). Laccase is essential for lignin degradation by the white-rot fungus Pycnoporus cinnabarinus. FEBS Lett 407, 89–92.[CrossRef]
    [Google Scholar]
  18. Erickson, T., Liu, L., Gueyikian, A., Zhu, X., Gibbons, J. & Williamson, P. R. ( 2001; ). Multiple virulence factors of Cryptococcus neoformans are dependent on VPH1. Mol Microbiol 42, 1121–1131.[CrossRef]
    [Google Scholar]
  19. Feldmesser, M., Rivera, J., Kress, Y., Kozel, T. R. & Casadevall, A. ( 2000; ). Antibody interactions with the capsule of Cryptococcus neoformans. Infect Immun 68, 3642–3650.[CrossRef]
    [Google Scholar]
  20. Garcia-Rivera, J. & Casadevall, A. ( 2001; ). Melanization of Cryptococcus neoformans reduces its susceptibility to the antimicrobial effects of silver nitrate. Med Mycol 39, 353–357.[CrossRef]
    [Google Scholar]
  21. Glatman-Freedman, A., Martin, J. M., Riska, P. F., Bloom, B. R. & Casadevall, A. ( 1996; ). Monoclonal antibodies to surface antigens of Mycobacterium tuberculosis and their use in a modified enzyme-linked immunosorbent spot assay for detection of mycobacteria. J Clin Microbiol 34, 2795–2802.
    [Google Scholar]
  22. Gómez, B. L., Nosanchuk, J. D., Diez, S., Youngchim, S., Aisen, P., Cano, L. E., Restrepo, A., Casadevall, A. & Hamilton, A. J. ( 2001; ). Detection of melanin-like pigments in the dimorphic fungal pathogen Paracoccidioides brasiliensis in vitro and during infection. Infect Immun 69, 5760–5767.[CrossRef]
    [Google Scholar]
  23. Hill, Z. H. ( 1992; ). The function of melanin or six blind people examine an elephant. BioEssays 14, 49–56.[CrossRef]
    [Google Scholar]
  24. Howard, R. J., Ferrari, M. A., Roach, D. H. & Money, N. P. ( 1991; ). Penetration of hard substrates by a fungus employing enormous turgor pressures. Proc Natl Acad Sci U S A 88, 11281–11284.[CrossRef]
    [Google Scholar]
  25. Huffnagle, G. B., Chen, G.-H., Curtis, J. L., McDonald, R. A., Strieter, R. M. & Toews, G. B. ( 1995; ). Down-regulation of the afferent phase of T cell-mediated pulmonary inflammation and immunity by a high melanin-producing strain of Cryptococcus neoformans. J Immunol 155, 3507–3516.
    [Google Scholar]
  26. Jahn, B., Koch, A., Schmidt, A., Wanner, G., Gehringer, H., Bhakdi, S. & Brakhage, A. A. ( 1997; ). Isolation and characterization of a pigmentless-conidium mutant of Aspergillus fumigatus with altered conidial surface and reduced virulence. Infect Immun 65, 5110–5117.
    [Google Scholar]
  27. Kopecka, M., Gabriel, M., Takeo, K., Yamaguchi, M., Svoboda, A., Ohkusu, M., Hata, K. & Yoshida, S. ( 2001; ). Microtubules and actin cytoskeleton in Cryptococcus neoformans compared with ascomycetous budding and fission yeasts. Eur J Cell Biol 80, 303–311.[CrossRef]
    [Google Scholar]
  28. Kwon-Chung, K. J., Polacheck, I. & Popkin, T. J. ( 1982; ). Melanin-lacking mutants of Cryptococcus neoformans and their virulence for mice. J Bacteriol 150, 1414–1421.
    [Google Scholar]
  29. Luberto, C., Toffaletti, D. L., Wills, E. A., Tucker, S. C., Casadevall, A., Perfect, J. R., Hannun, Y. A. & Del Poeta, M. M. ( 2001; ). Roles for inositol-phosphoryl ceramide synthase 1 (IPC1) in pathogenesis of C. neoformans. Genes Dev 15, 201–212.[CrossRef]
    [Google Scholar]
  30. Luther, J. P. & Lipke, H. ( 1980; ). Degradation of melanin by Aspergillus fumigatus. Appl Environ Microbiol 40, 145–155.
    [Google Scholar]
  31. McFadden, D. C. & Casadevall, A. ( 2001; ). Capsule and melanin synthesis in Cryptococcus neoformans. Med Mycol 39, 19–30.[CrossRef]
    [Google Scholar]
  32. Money, N. P. ( 1997; ). Mechanism linking cellular pigmentation and pathogenicity in rice blast disease. Fungal Genet Biol 22, 151–152.[CrossRef]
    [Google Scholar]
  33. Money, N. P., Caesar-TonThat, T. C., Frederick, B. & Henson, J. M. ( 1998; ). Melanin synthesis is associated with changes in hyphopodial turgor, permeability, and wall rigidity in Gaeumannomyces graminis var. graminis. Fungal Genet Biol 24, 240–251.[CrossRef]
    [Google Scholar]
  34. Nosanchuk, J. D., Valadon, P., Feldmesser, M. & Casadevall, A. (1999a; ). Melanization of Cryptococcus neoformans in murine infection. Mol Cell Biol 19, 745–750.
    [Google Scholar]
  35. Nosanchuk, J. D., Rudolph, J., Rosas, A. L. & Casadevall, A. ( 1999b; ). Evidence that Cryptococcus neoformans is melanized in pigeon excreta: implications for pathogenesis. Infect Immun 67, 5477–5479.
    [Google Scholar]
  36. Nosanchuk, J. D., Rosas, A. L., Lee, S. C. & Casadevall, A. ( 2000; ). Melanisation of Cryptococcus neoformans in human brain tissue. Lancet 355, 2049–2050.[CrossRef]
    [Google Scholar]
  37. Nosanchuk, J. D., Ovalle, R. & Casadevall, A. ( 2001; ). Glyphosate inhibits melanization of Cryptococcus neoformans and prolongs survival of mice after systemic infection. J Infect Dis 183, 1093–1099.[CrossRef]
    [Google Scholar]
  38. Osumi, M. ( 1998; ). The ultrastructure of yeast: cell wall structure and formation. Micron 29, 207–233.[CrossRef]
    [Google Scholar]
  39. Pierini, L. M. & Doering, T. L. ( 2001; ). Spatial and temporal sequence of capsule construction in Cryptococcus neoformans. Mol Microbiol 41, 105–115.[CrossRef]
    [Google Scholar]
  40. Polacheck, I., Hearing, V. J. & Kwon-Chung, K. J. ( 1982; ). Biochemical studies of phenoloxidase and utilization of catecholamines in Cryptococcus neoformans. J Bacteriol 150, 1212–1220.
    [Google Scholar]
  41. Ratto, M., Chatani, M., Ritschkoff, A. C. & Viikari, L. ( 2001; ). Screening of micro-organisms for decolorization of melanins produced by bluestain fungi. Appl Microbiol Biotechnol 55, 210–213.[CrossRef]
    [Google Scholar]
  42. Rhodes, J. C., Polacheck, I. & Kwon-Chung, K. J. ( 1982; ). Phenoloxidase activity and virulence in isogenic strains of Cryptococcus neoformans. Infect Immun 36, 1175–1184.
    [Google Scholar]
  43. Rosas, A. L. & Casadevall, A. ( 1997; ). Melanization affects susceptibility of Cryptococcus neoformans to heat and cold. FEMS Microbiol Lett 153, 265–272.[CrossRef]
    [Google Scholar]
  44. Rosas, A. L., Nosanchuk, J. D., Gomez, B. L., Edens, W. A., Henson, J. M. & Casadevall, A. ( 2000a; ). Isolation and serological analyses of fungal melanins. J Immunol Methods 244, 69–80.[CrossRef]
    [Google Scholar]
  45. Rosas, A. L., Nosanchuk, J. D., Feldmesser, M., Cox, G. M., McDade, H. C. & Casadevall, A. ( 2000b; ). Synthesis of polymerized melanin by Cryptococcus neoformans in infected rodents. Infect Immun 68, 2845–2853.[CrossRef]
    [Google Scholar]
  46. Rosas, A. L., Nosanchuk, J. D. & Casadevall, A. ( 2001; ). Passive immunization with melanin-binding monoclonal antibodies prolongs survival of mice with lethal Cryptococcus neoformans infection. Infect Immun 69, 3410–3412.[CrossRef]
    [Google Scholar]
  47. Rosas, A. L., MacGill, R. S., Nosanchuk, J. D., Kozel, T. R. & Casadevall, A. ( 2002; ). Activation of the alternative complement pathway by fungal melanins. Clin Diagn Lab Immunol 9, 144–148.
    [Google Scholar]
  48. Sekiya-Kawasaki, M., Abe, M., Saka, A., Watanabe, D., Kono, K., Minemura-Asakawa, M., Ishihara, S., Watanabe, T. & Ohya, Y. ( 2002; ). Dissection of upstream regulatory components of the Rho1p effector, 1,3-beta-glucan synthase, in Saccharomyces cerevisiae. Genetics 162, 663–676.
    [Google Scholar]
  49. Staib, F. ( 1962; ). Cryptococcus neoformans and Guizotia abyssicnica (syn. G. oleifera D.C.). Z Hyg 148, 466–475.[CrossRef]
    [Google Scholar]
  50. Staib, F. ( 1963; ). New concepts in the occurrence and identification of Cryptococcus neoformans. Mycopathol Mycol Appl 19, 143–145.
    [Google Scholar]
  51. Steenbergen, J. N., Shuman, H. A. & Casadevall, A. ( 2001; ). Cryptococcus neoformans interactions with amoebae suggest an explanation for its virulence and intracellular pathogenic strategy in macrophages. Proc Natl Acad Sci U S A 11, 11.
    [Google Scholar]
  52. Tsai, H. F., Fujii, I., Watanabe, A., Wheeler, M. H., Chang, Y. C., Yasuoka, Y., Ebizuka, Y. & Kwon-Chung, K. J. ( 2001; ). Pentaketide melanin biosynthesis in Aspergillus fumigatus requires chain-length shortening of a heptaketide precursor. J Biol Chem 276, 29292–29298.[CrossRef]
    [Google Scholar]
  53. Van Duin, D., Casadevall, A. & Nosanchuk, J. D. ( 2002; ). Melanization of Cryptococcus neoformans and Histoplasma capsulatum reduces their susceptibility to amphotericin B and caspofungin. Antimicrob Agents Chemother 46, 3394–3400.[CrossRef]
    [Google Scholar]
  54. Wang, Y. & Casadevall, A. ( 1994a; ). Decreased susceptibility of melanized Cryptococus neoformans to the fungicidal effects of ultraviolet light. Appl Environ Microbiol 60, 3864–3866.
    [Google Scholar]
  55. Wang, Y. & Casadevall, A. ( 1994b; ). Susceptibility of melanized and nonmelanized Cryptococcus neoformans to nitrogen- and oxygen-derived oxidants. Infect Immun 62, 3004–3007.
    [Google Scholar]
  56. Wang, Y. & Casadevall, A. ( 1996; ). Melanin, melanin ‘ghosts’, and melanin composition in Cryptococcus neoformans. Infect Immun 64, 2420–2424.
    [Google Scholar]
  57. Wang, Y., Aisen, P. & Casadevall, A. ( 1995; ). Cryptococcus neoformans melanin and virulence: mechanism of action. Infect Immun 63, 3131–3136.
    [Google Scholar]
  58. Wheeler, M. H. & Bell, A. A. ( 1988; ). Melanins and their importance in pathogenic fungi. Curr Topics Med Mycol 2, 338–387.
    [Google Scholar]
  59. Williamson, P. R. ( 1994; ). Biochemical and molecular characterization of the diphenol oxidase of Cryptococcus neoformans: identification as a laccase. J Bacteriol 176, 656–664.
    [Google Scholar]
  60. Zhu, X., Gibbons, J., Garcia-Rivera, J., Casadevall, A. & Williamson, P. R. ( 2001; ). Laccase of Cryptococcus neoformans is a cell wall-associated virulence factor. Infect Immun 69, 5589–5596.[CrossRef]
    [Google Scholar]
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