Considerable progress has been made in the selection and characterization of mutants that are defective in the synthesis of ergosterol in the yeast, Saccharomyces cerevisiae. Mutations in nearly every step of the yeast sterol biosynthetic pathway have been induced and selected. These mutants have been used to elucidate the sequential order of steps in sterol synthesis, to study the mode of action of antifungal agents and to determine the method of resistance of some pathogenic fungi, and to answer questions on the role of sterols in general cell biology. Physiological examination of ergosterol null mutants, lacking all biochemical activity attributed to the particular gene, supports a role for ergosterol in a number of critical functions in the organism. Among the physiological functions attributed to ergosterol are sparking and bulking requirements, involvement in amino acid and pyrimidine transport, resistance to antifungal agents and certain cations, and a requirement for respiratory activity. Those genetic null alleles discussed in this review are erg24, lacking the ability to reduce the delta 14 double bond; erg6, unable to methylate C-24; and erg3, defective in the C-5 desaturase. The different biochemical activities that are disrupted in the ergosterol mutants support a role for ergosterol in a number of critical functions in yeast.