In , glycosylphosphatidylinositol (GPI)-anchored cell wall mannoproteins, including -agglutinin, are secreted to the cell surface through vesicular transport pathways. At the cell surface the GPI anchors are cleaved within the glycan, then transglycosylated to form a covalent cross-link to 1,6--glucan. Among mutants that were temperature-sensitive for growth and for ability to cross-link the mannoprotein -agglutinin to the cell wall, one strain was complemented by , which encodes an ER-Golgi v-SNARE. Temperature-sensitive mutations in caused aberrations in cell wall structure, including excretion of -agglutinin into the medium, sensitivity to lysis with Zymolyase and hypersensitivity to Calcofluor White. At restrictive temperatures, mutations block secretion of invertase and other proteins, but -agglutinin was excreted into the extracellular medium. In wild-type parental or cells, secretion of -agglutinin also continued after protein synthesis was blocked with cycloheximide. This secretion was due to continued export of a significant amount of -agglutinin from compartments distal to the -dependent secretion step. Thus, in cells the ER-Golgi block allowed secretion to continue, but prevented cell wall incorporation of the -agglutinin. Therefore, a mutation early in the secretion pathway caused aberrant cell wall synthesis by preventing localization of key components required in wall cross-links.


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