Characterization of the CDC10 product and the timing of events of the budding site of Saccharomyces cerevisiae

Budding cells of the yeast Saccharomyces cerevisiae possess a ring of septin filaments of unknown biochemical nature that lies under the inner surface of the plasma membrane in the neck that connects the mother cell to its bud. Mutants, defective in any of the four genes (CDC3, CDC10, CDC11, CDC12), lack these septin filaments and display a pleiotropic phenotype that involves abnormal bud growth and an inability to complete cytokinesis. The cloned CDC10 was fused to bacterial genes to generate antibodies specific for the CDC10 product was a constituent of the septin filaments. Cdc10p-specific antibodies for septin staining and actin-specific rhodamine-phalloidine were used to investigate the timing of the localization of septin and actin at the budding site using the immunofluorescence microscopic technique. In wild-type cells, the timing of the appearance and disappearance of these proteins was indistinguishable. In addition, the cdc10 mutant did not prevent actin localization at the budding site. The mutant that was blocked in the actin function also did not prevent the septin localization of the Cdc10p. This result may suggest an organizational independence between these proteins in the bud formation. Finally, the localization of septin and actin in the cdc24 mutant cell was examined. It was found that the CDC24 function was necessary for the organization of septin and actin at the budding site.