Surface hydrophobicity changes of two Candida albicans serotype B mnn4delta mutants.

Department of Pathology, University of Virginia Health System, Charlottesville, Virginia 22908-0904, USA.
Eukaryotic Cell (Impact Factor: 3.59). 05/2005; 4(4):639-48. DOI: 10.1128/EC.4.4.639-648.2005
Source: PubMed

ABSTRACT Cell surface hydrophobicity (CSH) of Candida species enhances virulence by promoting adhesion to host tissues. Biochemical analysis of yeast cell walls has demonstrated that the most significant differences between hydrophobic and hydrophilic yeasts are found in the acid-labile fraction of Candida albicans phosphomannoprotein, suggesting that this fraction is important in the regulation of the CSH phenotype. The acid-labile fraction of C. albicans is unique among fungi, in that it is composed of an extended polymer of beta-1,2-mannose linked to the acid-stable region of the N-glycan by a phosphodiester bond. C. albicans serotype A and B strains both contain a beta-1,2-mannose acid-labile moiety, but only serotype A strains contain additional beta-1,2-mannose in the acid-stable region. A knockout of the C. albicans homolog of the Saccharomyces cerevisiae MNN4 gene was generated in two serotype B C. albicans patient isolates by using homologous gene replacement techniques, with the anticipation that they would be deficient in the acid-labile fraction and, therefore, demonstrate perturbed CSH. The resulting mnn4delta-deficient derivative has no detectable phosphate-linked beta-1,2-mannose in its cell wall, and hydrophobicity is increased significantly under conditions that promote the hydrophilic phenotype. The mnn4delta mutant also demonstrates an unanticipated perturbation in the acid-stable mannan fraction. The present study reports the first genetic knockout constructed in a serotype B C. albicans strain and represents an important step for dissecting the regulation of CSH.

  • [Show abstract] [Hide abstract]
    ABSTRACT: The cell wall proteins of fungi are modified by N- and O-linked mannosylation and phosphomannosylation, resulting in changes to the physical and immunological properties of the cell. Glycosylation of cell wall proteins involves the activities of families of endoplasmic reticulum and Golgi-located glycosyl transferases whose activities are difficult to infer through bioinformatics. The Candida albicans MNT1/KRE2 mannosyl transferase family is represented by five members. We showed previously that Mnt1 and Mnt2 are involved in O-linked mannosylation and are required for virulence. Here, the role of C. albicans MNT3, MNT4, and MNT5 was determined by generating single and multiple MnTDelta null mutants and by functional complementation experiments in Saccharomyces cerevisiae. CaMnt3, CaMnt4, and CaMnt5 did not participate in O-linked mannosylation, but CaMnt3 and CaMnt5 had redundant activities in phosphomannosylation and were responsible for attachment of approximately half of the phosphomannan attached to N-linked mannans. CaMnt4 and CaMnt5 participated in N-mannan branching. Deletion of CaMNT3, CaMNT4, and CaMNT5 affected the growth rate and virulence of C. albicans, affected the recognition of the yeast by human monocytes and cytokine stimulation, and led to increased cell wall chitin content and exposure of beta-glucan at the cell wall surface. Therefore, the MNT1/KRE2 gene family participates in three types of protein mannosylation in C. albicans, and these modifications play vital roles in fungal cell wall structure and cell surface recognition by the innate immune system.
    Journal of Biological Chemistry 02/2010; 285(16):12087-95. · 4.65 Impact Factor
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Glycosyltransferases are specific enzymes that catalyse the transfer of monosaccharide moieties to biological substrates, including proteins, lipids and carbohydrates. These enzymes are present from prokaryotes to humans, and their glycoconjugate products are often vital for survival of the organism. Many glycosyltransferases found in fungal pathogens such as Cryptococcus neoformans do not exist in mammalian systems, making them attractive potential targets for selectively toxic agents. In this article, we present the features of this diverse class of enzymes, and review the fungal glycosyltransferases that are involved in synthesis of the cell wall, the cryptococcal capsule, glycoproteins and glycolipids. We specifically focus on enzymes that have been identified or studied in C. neoformans, and we consider future directions for research on glycosyltransferases in the context of this opportunistic pathogen.
    FEMS Yeast Research 07/2006; 6(4):499-512. · 2.46 Impact Factor
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: The aim of the present paper is to evaluate the effect of the high molecular weight chitosan (HMWC) and of sodium alginate (NaAL) on surface hydrophobicity of Candida albicans and on adhesion of the yeast to epithelial cells and fibroblasts of different proceeding. For this study, a collection strain and seven isolates of C. albicans from saliva (patients with denture stomatitis) were grown in Sabouraud glucose agar supplemented with HMWC or NaAL or in absence of them (control). Hydrophobicity was determined by adhesion to hydrocarbons method using two organic media (xylene and chloroform). For adhesion experiments, aqueous suspensions of yeasts were contacted with solutions of biopolymers and different cells (rat and human fibroblasts and epithelial cells Hep-2). The quantification of adhesion was made by optical microscopy. RESULTS: A decrease in hydrophobicity was observed in the presence of HMWC (44%) and of NaAL (82%) when chloroform was employed as organic medium, meanwhile the decreases were of 30% with HMWC and 19% with NaAL in the presence of xylene. Adhesion of C. albicans to epithelial cells and human fibroblasts decreased significantly with both biopolymers. In the case of rat fibroblasts, a decrease was observed only with NaAL. None of experiments showed significant differences associated to fibroblast type. CONCLUSIONS: Biopolymers showed effectiveness in reducing hydrophobicity and adhesion of C. albicans to cells, which are important virulence factors related to colonization of the soft tissues of host or acrylic surfaces present in the oral system.
    Medicina oral, patologia oral y cirugia bucal 04/2006; 11(2):E120-5. · 1.02 Impact Factor