Abstract The sequence of posttranslational events in the export of yeast glycoproteins has been determined with the aid of mutants that affect the secretory apparatus. Temperature-sensitive secretory mutants ( sec ) of S. cerevisiae, when incubated at a nonpermissive growth temperature (37°C), accumulate intracellular precursor forms of exported glycoproteins, such as invertase, and expand or amplify one or more of three different secretory organelles. Characterization of haploid double- sec -mutant strains, with regard to the structure of the accumulated invertase and the morphology of the exaggerated organelles, allows assessment of the order in which the gene products are required, the sequence of invertase maturation steps and a pathway of secretory organelles. The transitions from one organelle to the next require energy and sec gene products. One of the mutants ( sec7 ) accumulates a different organelle depending on the concentration of glucose in the medium. In normal growth medium (2% glucose), a thermally irreversible structure, the Berkeley body, predominates; in low glucose (0.1%), Golgi structures accumulate thermoreversibly. The results are consistent with the following model. Secretory proteins enter the ER, where the initial steps of glycosylation occur. Nine or more sec gene products and energy are required to transfer material to a Golgi-like structure, where further glycosylation occurs. Two or more functions and energy are required to package nearly fully glycosylated proteins into vesicles that are then transported into the bud, where they fuse with the plasma membrane in a process that requires at least ten additional gene products and energy.