Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1994 Apr;176(8):2318–2325. doi: 10.1128/jb.176.8.2318-2325.1994

Isolation and characterization of yeast monomorphic mutants of Candida albicans.

M V Elorza 1, R Sentandreu 1, J Ruiz-Herrera 1
PMCID: PMC205355  PMID: 8157600

Abstract

A method was devised for the isolation of yeast monomorphic (LEV) mutants of Candida albicans. By this procedure, about 20 stable yeast-like mutants were isolated after mutagenesis with ethyl methane sulfonate. The growth rate of the mutants in different carbon sources, both fermentable and not, was indistinguishable from that of the parental strain, but they were unable to grow as mycelial forms after application of any of the common effective inducers, i.e., heat shock, pH alterations, proline addition, or use of GlcNAc as the carbon source. Studies performed with one selected strain demonstrated that it had severe alterations in the chemical composition of the cell wall, mainly in the levels of chitin and glucans, and in specific mannoproteins, some of them recognizable by specific polyclonal and monoclonal antibodies. It is suggested that these structural alterations hinder the construction of a normal hyphal wall.

Full text

PDF
2318

Images in this article

2319Image
on p.2319
2322Image
on p.2322
2323Image
on p.2323
2323Image
on p.2323
2324Image
on p.2324

Selected References

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

  1. Anderson J. M., Soll D. R. Unique phenotype of opaque cells in the white-opaque transition of Candida albicans. J Bacteriol. 1987 Dec;169(12):5579–5588. doi: 10.1128/jb.169.12.5579-5588.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bartnicki-Garcia S., Lippman E. Fungal morphogenesis: cell wall construction in Mucor rouxii. Science. 1969 Jul 18;165(3890):302–304. doi: 10.1126/science.165.3890.302. [DOI] [PubMed] [Google Scholar]
  3. Burnette W. N. "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem. 1981 Apr;112(2):195–203. doi: 10.1016/0003-2697(81)90281-5. [DOI] [PubMed] [Google Scholar]
  4. Cano-Canchola C., Sosa L., Fonzi W., Sypherd P., Ruiz-Herrera J. Developmental regulation of CUP gene expression through DNA methylation in Mucor spp. J Bacteriol. 1992 Jan;174(2):362–366. doi: 10.1128/jb.174.2.362-366.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cano C., Herrera-Estrella L., Ruiz-Herrera J. DNA methylation and polyamines in regulation of development of the fungus Mucor rouxii. J Bacteriol. 1988 Dec;170(12):5946–5948. doi: 10.1128/jb.170.12.5946-5948.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Casanova M., Gil M. L., Cardeñoso L., Martinez J. P., Sentandreu R. Identification of wall-specific antigens synthesized during germ tube formation by Candida albicans. Infect Immun. 1989 Jan;57(1):262–271. doi: 10.1128/iai.57.1.262-271.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Casanova M., Martínez J. P., Chaffin W. L. Fab fragments from a monoclonal antibody against a germ tube mannoprotein block the yeast-to-mycelium transition in Candida albicans. Infect Immun. 1990 Nov;58(11):3810–3812. doi: 10.1128/iai.58.11.3810-3812.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Chattaway F. W., Holmes M. R., Barlow A. J. Cell wall composition of the mycelial and blastospore forms of Candida albicans. J Gen Microbiol. 1968 May;51(3):367–376. doi: 10.1099/00221287-51-3-367. [DOI] [PubMed] [Google Scholar]
  9. Chen-Wu J. L., Zwicker J., Bowen A. R., Robbins P. W. Expression of chitin synthase genes during yeast and hyphal growth phases of Candida albicans. Mol Microbiol. 1992 Feb;6(4):497–502. doi: 10.1111/j.1365-2958.1992.tb01494.x. [DOI] [PubMed] [Google Scholar]
  10. Dabrowa N., Taxer S. S., Howard D. H. Germination of Candida albicans induced by proline. Infect Immun. 1976 Mar;13(3):830–835. doi: 10.1128/iai.13.3.830-835.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Elorza M. V., Murgui A., Sentandreu R. Dimorphism in Candida albicans: contribution of mannoproteins to the architecture of yeast and mycelial cell walls. J Gen Microbiol. 1985 Sep;131(9):2209–2216. doi: 10.1099/00221287-131-9-2209. [DOI] [PubMed] [Google Scholar]
  12. Elorza M. V., Sentandreu R. Effect of cycloheximide on yeast cell wall synthesis. Biochem Biophys Res Commun. 1969 Aug 22;36(5):741–747. doi: 10.1016/0006-291x(69)90672-x. [DOI] [PubMed] [Google Scholar]
  13. Elorza V., Mormeneo S., Garcia de la Cruz F., Gimeno C., Sentandreu R. Evidence for the formation of covalent bonds between macromolecules in the domain of the wall of Candida albicans mycelial cells. Biochem Biophys Res Commun. 1989 Aug 15;162(3):1118–1125. doi: 10.1016/0006-291x(89)90789-4. [DOI] [PubMed] [Google Scholar]
  14. Kennedy M. J., Rogers A. L., Hanselmen L. R., Soll D. R., Yancey R. J., Jr Variation in adhesion and cell surface hydrophobicity in Candida albicans white and opaque phenotypes. Mycopathologia. 1988 Jun;102(3):149–156. doi: 10.1007/BF00437397. [DOI] [PubMed] [Google Scholar]
  15. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. Marcilla A., Elorza M. V., Mormeneo S., Rico H., Sentandreu R. Candida albicans mycelial wall structure: supramolecular complexes released by zymolyase, chitinase and beta-mercaptoethanol. Arch Microbiol. 1991;155(4):312–319. doi: 10.1007/BF00243448. [DOI] [PubMed] [Google Scholar]
  18. Marcilla A., Mormeneo S., Elorza M. V., Manclus J. J., Sentandreu R. Wall formation by Candida albicans yeast cells: synthesis, secretion and incorporation of two types of mannoproteins. J Gen Microbiol. 1993 Dec;139(12):2985–2993. doi: 10.1099/00221287-139-12-2985. [DOI] [PubMed] [Google Scholar]
  19. Martinez J. P., Gil M. L., Casanova M., Lopez-Ribot J. L., Garcia De Lomas J., Sentandreu R. Wall mannoproteins in cells from colonial phenotypic variants of Candida albicans. J Gen Microbiol. 1990 Dec;136(12):2421–2432. doi: 10.1099/00221287-136-12-2421. [DOI] [PubMed] [Google Scholar]
  20. Martinez J. P., Lopez-Ribot J. L., Gil M. L., Sentandreu R., Ruiz-Herrera J. Inhibition of the dimorphic transition of Candida albicans by the ornithine decarboxylase inhibitor 1,4-diaminobutanone: alterations in the glycoprotein composition of the cell wall. J Gen Microbiol. 1990 Oct;136(10):1937–1943. doi: 10.1099/00221287-136-10-1937. [DOI] [PubMed] [Google Scholar]
  21. Mattia E., Carruba G., Angiolella L., Cassone A. Induction of germ tube formation by N-acetyl-D-glucosamine in Candida albicans: uptake of inducer and germinative response. J Bacteriol. 1982 Nov;152(2):555–562. doi: 10.1128/jb.152.2.555-562.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Merz W. G., Connelly C., Hieter P. Variation of electrophoretic karyotypes among clinical isolates of Candida albicans. J Clin Microbiol. 1988 May;26(5):842–845. doi: 10.1128/jcm.26.5.842-845.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Murgui A., Elorza M. V., Sentandreu R. Effect of papulacandin B and calcofluor white on the incorporation of mannoproteins in the wall of Candida albicans blastospores. Biochim Biophys Acta. 1985 Aug 16;841(2):215–222. doi: 10.1016/0304-4165(85)90024-8. [DOI] [PubMed] [Google Scholar]
  24. Murgui A., Elorza M. V., Sentandreu R. Tunicamycin and papulacandin B inhibit incorporation of specific mannoproteins into the wall of Candida albicans regenerating protoplasts. Biochim Biophys Acta. 1986 Dec 10;884(3):550–558. doi: 10.1016/0304-4165(86)90207-2. [DOI] [PubMed] [Google Scholar]
  25. Rustchenko-Bulgac E. P., Howard D. H. Multiple chromosomal and phenotypic changes in spontaneous mutants of Candida albicans. J Gen Microbiol. 1993 Jun;139(Pt 6):1195–1207. doi: 10.1099/00221287-139-6-1195. [DOI] [PubMed] [Google Scholar]
  26. Rustchenko-Bulgac E. P., Sherman F., Hicks J. B. Chromosomal rearrangements associated with morphological mutants provide a means for genetic variation of Candida albicans. J Bacteriol. 1990 Mar;172(3):1276–1283. doi: 10.1128/jb.172.3.1276-1283.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. San-Blas F., San-Blas G., Cova L. J. A morphological mutant of Paracoccidioides brasiliensis strain IVIC Pb9. Isolation and wall characterization. J Gen Microbiol. 1976 Apr;93(2):209–218. doi: 10.1099/00221287-93-2-209. [DOI] [PubMed] [Google Scholar]
  28. Simonetti N., Strippoli V., Cassone A. Yeast-mycelial conversion induced by N-acetyl-D-glucosamine in Candida albicans. Nature. 1974 Jul 26;250(464):344–346. doi: 10.1038/250344a0. [DOI] [PubMed] [Google Scholar]
  29. Slutsky B., Buffo J., Soll D. R. High-frequency switching of colony morphology in Candida albicans. Science. 1985 Nov 8;230(4726):666–669. doi: 10.1126/science.3901258. [DOI] [PubMed] [Google Scholar]
  30. Sobel J. D., Muller G., Buckley H. R. Critical role of germ tube formation in the pathogenesis of candidal vaginitis. Infect Immun. 1984 Jun;44(3):576–580. doi: 10.1128/iai.44.3.576-580.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Soll D. R., Herman M. A., Staebell M. A. The involvement of cell wall expansion in the two modes of mycelium formation of Candida albicans. J Gen Microbiol. 1985 Sep;131(9):2367–2375. doi: 10.1099/00221287-131-9-2367. [DOI] [PubMed] [Google Scholar]
  32. Suzuki T., Kobayashi I., Kanbe T., Tanaka K. High frequency variation of colony morphology and chromosome reorganization in the pathogenic yeast Candida albicans. J Gen Microbiol. 1989 Feb;135(Pt 2):425–434. doi: 10.1099/00221287-135-2-425. [DOI] [PubMed] [Google Scholar]
  33. Suzuki T., Miyamae Y., Ishida I. Variation of colony morphology and chromosomal rearrangement in Candida tropicalis pK233. J Gen Microbiol. 1991 Jan;137(1):161–167. doi: 10.1099/00221287-137-1-161. [DOI] [PubMed] [Google Scholar]
  34. Torosantucci A., Gomez M. J., Bromuro C., Casalinuovo I., Cassone A. Biochemical and antigenic characterization of mannoprotein constituents released from yeast and mycelial forms of Candida albicans. J Med Vet Mycol. 1991;29(6):361–372. doi: 10.1080/02681219180000591. [DOI] [PubMed] [Google Scholar]
  35. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Wain W. H., Brayton A. R., Cawson R. A. Variations in the response to N-acetyl-D-glucosamine by isolates of Candida albicans. Mycopathologia. 1976 Jun 4;58(1):27–29. doi: 10.1007/BF00493590. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES