Schizosaccharomyces pombe Map4 adhesin is a glycoprotein that can be extracted from the cell wall with alkali but not with β-glucanases and requires the C-terminal DIPSY domain for function

• Published in: Molecular microbiology 2008-09, Vol.69 (6), p.1476-1490
• Main Authors: Sharifmoghadam, Mohammad Reza, Valdivieso, M.-Henar
• Format: Article
• Language: English
• Subjects: Biological and medical sciences; Cell Adhesion; Cell Adhesion Molecules - chemistry; Cell Adhesion Molecules - genetics; Cell Adhesion Molecules - isolation & purification; Cell Wall - chemistry; Fundamental and applied biological sciences. Psychology; Glycoproteins - chemistry; Glycoproteins - genetics; Glycoproteins - isolation & purification; Microbiology; Miscellaneous; Mycology; Protein Structure, Tertiary; Schizosaccharomyces - chemistry; Schizosaccharomyces - physiology; Schizosaccharomyces pombe Proteins - chemistry; Schizosaccharomyces pombe Proteins - genetics; Schizosaccharomyces pombe Proteins - isolation & purification
• ISSN: 0950-382X; 1365-2958
• DOI: 10.1111/j.1365-2958.2008.06375.x

In fungi, cell adhesion is required for flocculation, mating and virulence, and it is mediated by covalently bound cell wall proteins termed adhesins. Map4, an adhesin required for mating in Schizosaccharomyces pombe, is N-glycosylated and O-glycosylated, and is an endogenous substrate for the mannosyl transferase Oma4p. Map4 has a modular structure with an N-terminal signal peptide, a serine and threonine (S/T)-rich domain that includes nine repeats of 36 amino acids (rich in serine and threonine residues, but lacking glutamines), and a C-terminal DIPSY domain with no glycosylphosphatidyl inositol (GPI)-anchor signal. Map4 can be extracted from cell walls with SDS/mercaptoethanol sample buffer or with mild alkali solutions. After extensive extraction with hot sample buffer, no more protein can be released by β-glucanases or alkali. Additionally, none of the cysteine residues of the protein is required for its retention at the cell wall. These results show that Map4 is not directly bound to β-glucans and point to the existence of alkali- and SDS/mercaptoethanol-sensitive linkages between cell wall proteins. The N-terminal S/T-rich regions are required for cell wall attachment, but the C-terminal DIPSY domain is required for agglutination and mating in liquid and solid media.