1887

Abstract

The homologues of the most studied stationary-phase genes, and , were used to test the hypothesis that, within a biofilm, some cells reach stationary phase within continuously fed, as well as static, biofilms grown on dental acrylic. The authors first studied the expression patterns of these two genes in planktonic growth conditions. Using real-time RT-PCR (RT-RTPCR), increased peak expression of both and was observed at 5 and 6 days, respectively, in grown in suspension culture. –yellow fluorescent protein (YFP) and were constructed to study expression at the cellular level and protein localization in . Snz1p–YFP and Sno1p–YFP localized to the cytoplasm with maximum expression (>90 %) at 5 and 6 days, respectively, in planktonic conditions. When yeast growth was reinitiated, loss of fluorescence began immediately. Germ tubes and hyphae were non-fluorescent. Pseudohyphae began appearing at 9 days in planktonic yeast culture and expressed each protein by 11 days; however, the cells budding from pseudohyphae were not fluorescent. Biofilm was formed under either static or continuously fed conditions. Increased expression of the two genes was shown by RT-RTPCR, beginning by day 3 and increasing through to day 15 (continuously fed biofilm). Only the bottommost layer of acrylic-adhered cells in the biofilm showed 25 and 40 % fluorescence at 6 and 15 days, respectively. These observations suggest that only a few cells in biofilms express genes associated with the planktonic stationary phase and that these are found at the bottom of the biofilm adhered to the surface.

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2006-07-01
2019-10-23
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References

  1. Adams, A., Gottschling, D. E., Kaiser, C. A. & Stearns, T. ( 1997; ). Methods in Yeast Geneticsa Cold Spring Harbor Laboratory Course Manual. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
  2. Altschul, S. F., Madden, T. L., Schaffer, A. A., Zhang, J., Zhang, Z., Miller, W. & Lipman, D. J. ( 1997; ). Gapped blast and psi-blast: a new generation of protein database search programs. Nucleic Acids Res 25, 3389–3402.[CrossRef]
    [Google Scholar]
  3. Arnaud, M. B., Costanzo, M. C., Skrzypek, M. S., Binkley, G., Lane, C., Miyasato, S. R. & Sherlock, G. ( 2005; ). The Candida genome database (CGD), a community resource for Candida albicans gene and protein information. Nucleic Acids Res 33, D358–D363.[CrossRef]
    [Google Scholar]
  4. Baillie, G. S. & Douglas, L. J. ( 1999; ). Role of dimorphism in the development of Candida albicans biofilms. J Med Microbiol 48, 671–679.[CrossRef]
    [Google Scholar]
  5. Baillie, G. S. & Douglas, L. J. ( 2000; ). Matrix polymers of Candida biofilms and their possible role in biofilm resistance to antifungal agents. J Antimicrob Chemother 46, 397–403.[CrossRef]
    [Google Scholar]
  6. Balakrishnan, R., Christie, K. R., Costanzo, M. C. & 24 other authors ( 2002; ). Saccharomyces Genome Database. http://www.yeastgenome.org/, visited 22 September 2002.
  7. Barelle, C. J., Bohula, E. A., Kron, S. J., Wessels, D., Soll, D. R., Schafer, A., Brown, A. J. & Gow, N. A. ( 2003; ). Asynchronous cell cycle and asymmetric vacuolar inheritance in true hyphae of Candida albicans. Eukaryot Cell 2, 398–410.[CrossRef]
    [Google Scholar]
  8. Braun, E. L., Fuge, E. K., Padilla, P. A. & Werner-Washburne, M. ( 1996; ). A stationary-phase gene in Saccharomyces cerevisiae is a member of a novel, highly conserved gene family. J Bacteriol 178, 6865–6872.
    [Google Scholar]
  9. Chandra, J., Kuhn, D. M., Mukherjee, P. K., Hoyer, L. L., McCormick, T. & Ghannoum, M. A. ( 2001; ). Biofilm formation by the fungal pathogen Candida albicans: development, architecture, and drug resistance. J Bacteriol 183, 5385–5394.[CrossRef]
    [Google Scholar]
  10. Douglas, L. J. ( 2003; ). Candida biofilms and their role in infection. Trends Microbiol 11, 30–36.[CrossRef]
    [Google Scholar]
  11. Gerami-Nejad, M., Berman, J. & Gale, C. A. ( 2001; ). Cassettes for PCR-mediated construction of green, yellow, and cyan fluorescent protein fusions in Candida albicans. Yeast 18, 859–864.[CrossRef]
    [Google Scholar]
  12. Huh, W. K., Falvo, J. V., Gerke, L. C., Carroll, A. S., Howson, R. W., Weissman, J. S. & O'Shea, E. K. ( 2003; ). Global analysis of protein localization in budding yeast. Nature 425, 686–691.[CrossRef]
    [Google Scholar]
  13. Kumamoto, C. A. & Vinces, M. D. ( 2005; ). Alternative Candida albicans lifestyles: growth on surfaces. Annu Rev Microbiol 59, 113–133.[CrossRef]
    [Google Scholar]
  14. Lamarre, C., LeMay, J. D., Deslauriers, N. & Bourbonnais, Y. ( 2001; ). Candida albicans expresses an unusual cytoplasmic manganese-containing superoxide dismutase (SOD3 gene product) upon the entry and during the stationary phase. J Biol Chem 276, 43784–43791.[CrossRef]
    [Google Scholar]
  15. Maneu, V., Martinez, J. P. & Gozalbo, D. ( 2000; ). Identification of Candida albicans clinical isolates by PCR amplification of an EFB1 gene fragment containing an intron-interrupted open reading frame. Med Mycol 38, 123–126.[CrossRef]
    [Google Scholar]
  16. Moreno, I., Pedreno, Y., Maicas, S., Sentandreu, R., Herrero, E. & Valentin, E. ( 2003; ). Characterization of a Candida albicans gene encoding a putative transcriptional factor required for cell wall integrity. FEMS Microbiol Lett 226, 159–167.[CrossRef]
    [Google Scholar]
  17. Mukherjee, P. K., Zhou, G., Munyon, R. & Ghannoum, M. A. ( 2005; ). Candida biofilm: a well-designed protected environment. Med Mycol 43, 191–208.[CrossRef]
    [Google Scholar]
  18. Radonjic, M., Andrau, J. C., Lijnzaad, P., Kemmeren, P., Kockelkorn, T. T., van Leenen, D., van Berkum, N. L. & Holstege, F. C. ( 2005; ). Genome-wide analyses reveal RNA polymerase II located upstream of genes poised for rapid response upon S. cerevisiae stationary phase exit. Mol Cell 18, 171–183.[CrossRef]
    [Google Scholar]
  19. Ramage, G., Vandewalle, K., Wickes, B. L. & Lopez-Ribot, J. L. ( 2001; ). Characteristics of biofilm formation by Candida albicans. Rev Iberoam Micol 18, 163–170.
    [Google Scholar]
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