Posttranslational modifications required for cell surface localization and function of the fungal adhesin Aga1p.
Adherence of fungal cells to host substrates and each other affects their access to nutrients, sexual conjugation, and survival in hosts. Adhesins are cell surface proteins that mediate these different cell adhesion interactions. In this study, we examine the in vivo functional requirements for specific posttranslational modifications to these proteins, including glycophosphatidylinositol (GPI) anchor addition and O-linked glycosylation. The processing of some fungal GPI anchors, creating links to cell wall beta-1,6 glucans, is postulated to facilitate postsecretory traffic of proteins to cell wall domains conducive to their functions. By studying the yeast sexual adhesin subunit Aga1p, we found that deletion of its signal sequence for GPI addition eliminated its activity, while deletions of different internal domains had various effects on function. Substitution of the Aga1p GPI signal domain with those of other GPI-anchored proteins, a single transmembrane domain, or a cysteine capable of forming a disulfide all produced functional adhesins. A portion of the cellular pool of Aga1p was determined to be cell wall resident. Aga1p and the alpha-agglutinin Agalpha1p were shown to be under glycosylated in cells lacking the protein mannosyltransferase genes PMT1 and PMT2, with phenotypes manifested only in MATalpha cells for single mutants but in both cell types when both genes are absent. We conclude that posttranslational modifications to Aga1p are necessary for its biogenesis and activity. Our studies also suggest that in addition to GPI-glucan linkages, other cell surface anchorage mechanisms, such as transmembrane domains or disulfides, may be employed by fungal species to localize adhesins.
Huang
Guohong
G
Department of Biology, Syracuse University, Syracuse, New York 13244, USA. ghuang@syr.edu
Zhang
Mingliang
M
Erdman
Scott E
SE
D000372:Agglutination Tests; D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D001483:Base Sequence; D015815:Cell Adhesion Molecules; D002462:Cell Membrane / Q000737:chemistry; D002473:Cell Wall / Q000737:chemistry; D017353:Gene Deletion; D005936:Glucans / Q000276:immunology / Q000502:physiology; D006031:Glycosylation; D017261:Glycosylphosphatidylinositols / Q000235:genetics / Q000502:physiology; D008364:Mannosyltransferases / Q000235:genetics / Q000502:physiology; D000072235:Mating Factor; D008565:Membrane Proteins / Q000235:genetics / Q000502:physiology; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional / Q000379:methods; D010455:Peptides / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D021382:Protein Sorting Signals / Q000235:genetics; D021381:Protein Transport; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000502:physiology; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000502:physiology*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000502:physiology*; D047071:beta-Glucans
C543384:AGA1 protein, S cerevisiae; D015815:Cell Adhesion Molecules; D005936:Glucans; D017261:Glycosylphosphatidylinositols; D008565:Membrane Proteins; D010455:Peptides; D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; D047071:beta-Glucans; C069939:a-agglutinin protein, S cerevisiae; C064197:beta-1,6-glucan; D000072235:Mating Factor; D008364:Mannosyltransferases; C082696:protein O-mannosyltransferase