The incorporation of mannoproteins in the cell wall of S. cerevisiae and filamentous Ascomycetes

In yeast, glucanase extractable cell wall proteins are anchored to the plasma membrane at an intermediate stage in their biogenesis via a glycosylphosphatidylinositol (GPI) moiety before they become anchored to the wall glucan via a β1,6-glucan linkage. The mechanism of the membrane processing step of cell wall proteins is not known. Here, we report that Ascomycete filamentous fungi involved in food spoilage such as Aspergillus, Paecilomyces and Penicillium, also contain GPI membrane-anchored proteins some of which are processed by an endogenous phospholipase C activity. Furthermore, similar to the situation in yeast, their cell walls contain mannoproteins which are linked to the glucan backbone through a β1,6-glucan linkage. Interestingly, one mould which contains a significant amount of non covalently linked β1,6-glucosylated cell wall proteins, is much more sensitive towards β1,3-glucanases and membrane perturbing peptides than the others.

[1]  F. Klis,et al.  Increased Cell Wall Porosity in Saccharomyces cerevisiae after Treatment with Dithiothreitol or EDTA , 1989 .

[2]  T. Nakajima,et al.  Chemical structure of the galactomannan moiety in the cell wall glycoproteins of Aspergillus oryzae , 1994 .

[3]  C. Bulawa.  Genetics and molecular biology of chitin synthesis in fungi. , 1993, Annual review of microbiology.

[4]  H. Bussey,et al.  Glycosyl phosphatidylinositol-dependent cross-linking of alpha- agglutinin and beta 1,6-glucan in the Saccharomyces cerevisiae cell wall , 1995, The Journal of cell biology.

[5]  S. Silverman,et al.  The function of chitin synthases 2 and 3 in the Saccharomyces cerevisiae cell cycle , 1991, The Journal of cell biology.

[6]  N. Fasel,et al.  Anchoring of an Immunogenic Plasmodium falciparum Circumsporozoite Protein on the Surface of Dictyostelium discoideum(*) , 1995, The Journal of Biological Chemistry.

[7]  F. Klis,et al.  The glucanase‐soluble mannoproteins limit cell wall porosity in Saccharomyces cerevisiae , 1990, Yeast.

[8]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[9]  J. C. Kapteyn,et al.  Glucosylation of cell wall proteins in regenerating spheroplasts of Candida albicans. , 1995, FEMS microbiology letters.

[10]  B. Brzobohatý,et al.  Factors enhancing genetic transformation of intact yeast cells modify cell wall porosity. , 1986, Journal of general microbiology.

[11]  Robert A. Samson,et al.  Introduction to food-borne fungi. , 1989 .

[12]  M. Snyder,et al.  Molecular basis of cell integrity and morphogenesis in Saccharomyces cerevisiae. , 1995, Microbiological reviews.

[13]  P. Lipke,et al.  A pathway for cell wall anchorage of Saccharomyces cerevisiae alpha-agglutinin , 1994, Molecular and cellular biology.

[14]  H. Towbin,et al.  Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. , 1979, Proceedings of the National Academy of Sciences of the United States of America.

[15]  F. Klis.  Review: Cell wall assembly in yeast , 1994, Yeast.

[16]  C. Ballou.  Isolation, characterization, and properties of Saccharomyces cerevisiae mnn mutants with nonconditional protein glycosylation defects. , 1990, Methods in enzymology.

[17]  F. Klis,et al.  Targeting of a heterologous protein to the cell wall of Saccharomyces cerevisiae , 1993, Yeast.

[18]  J. C. Kapteyn,et al.  Covalent association of beta-1,3-glucan with beta-1,6-glucosylated mannoproteins in cell walls of Candida albicans , 1995, Journal of bacteriology.

[19]  D. Wessel,et al.  A method for the quantitative recovery of protein in dilute solution in the presence of detergents and lipids. , 1984, Analytical biochemistry.

[20]  R. Dwek,et al.  Characterization of the cross-reacting determinant (CRD) of the glycosyl-phosphatidylinositol membrane anchor of Trypanosoma brucei variant surface glycoprotein. , 1988, European journal of biochemistry.

[21]  F. Klis,et al.  Glucosylation of fungal cell wall proteins as a potential target for novel antifungal agents , 1996 .

[22]  F. Klis,et al.  The retention mechanism of cell wall proteins in Saccharomyces cerevisiae. Wall-bound Cwp2p is beta-1,6-glucosylated. , 1996, Biochimica et biophysica acta.

[23]  J. C. Kapteyn,et al.  Identification of beta-1,6-glucosylated cell wall proteins in yeast and hyphal forms of Candida albicans. , 1994, European journal of cell biology.

[24]  G. Müller,et al.  Glucose-induced sequential processing of a glycosyl-phosphatidylinositol-anchored ectoprotein in Saccharomyces cerevisiae , 1996, Molecular and cellular biology.