Mushroom bodies in the Drosophila brain are centers for olfactory learning and memory. We have previously shown that the mushroom bodies comprise three types of neurons with distinct axonal projections. These three types of neurons are generated sequentially from common neuroblasts. We report here the identification of a gene that we have named enoki mushroom (enok), which when it is mutated gives rise to mushroom bodies with reduced axonal structures. enok encodes a putative histone acetyltransferase (HAT) of the MYST family, members of which have been implicated as important modulators of transcriptional activity. A single amino acid change in the zinc finger motif of the putative catalytic HAT domain gives the same phenotype as a null allele, and this finding indicates the importance of HAT activity to Enok's function. Further phenotypic analysis demonstrates that the mushroom body defect is due to an arrest in neuroblast proliferation rather than a failure of either cell fate switching or axon branching. Clonal analyses in the wing discs and the ovaries suggest that enok is essential for normal cell proliferation in some, but not all, tissues. Our results provide in vivo evidence for essential functions of a histone acetyltransferase in the construction of the Drosophila brain.