A meiotic fine structure map of the gal4 locus was constructed, which extended over 0.44 units on the chromosome (units in percent frequency of supposed recombination). Several nonsense gal4 mutations (four UAA and two supposed UGA [gal4-62 and gal4-69]) were placed at various sites on the map. In reversion experiments with 20 independently isolated gal4 mutants, only the gal4-62 and gal4-69 alleles, which are located at the same site on the map, could revert to overcome the superrepression of gal80s-1 spontaneously with a frequency of approximately 4 x 10(-7). Secondary mutations in the revertants occurred in the region of gal4-62 or were due to unlinked suppressors. A total of 15 GAL81 mutations in 19 isolates were found to be located in the same region as gal4-62 by three-point crosses with the aid of gal4 mutants; the other four could not be analyzed. The reverted gal4 gene and GAL81 mutations were semidominant over the wild-type GAL4+ allele and fully dominant over a nonsense gal4 mutation. Four suppressors (one dominant and three recessive) effective against gal4-62 and gal4-69 were isolated. The dominant suppressor was also effective against three independent, authentic auxotrophic UGA nonsense mutations, and one of the three recessive suppressors were effective against the authentic auxotrophic UAA and UAG mutations. These results strongly support the idea that the gal4 locus is expressed constitutively and codes for a regulatory protein. The GAL81 site mapped inside the locus codes for a part of the gal4 protein but does not work as an operator.