Aggregation of Dictyostelium discoideum amoebae into multicellular structures is organized by cyclic AMP (cAMP), which acts as a chemoattractant, as a second messenger, and as a morphogen. Aggregation of D. discoideum cells depends on the expression of adenylyl cyclase ACA, which provides extracellular cAMP for signal relay and intracellular cAMP for the induction of genes required at multicellular stages. We have identified a DNA-binding activity specific for a highly A+T-enriched motif in the upstream region of the ACA-encoding gene, acaA. The factor shows DNA-binding characteristics very similar to those of C-module-binding factor (CbfA). Although CbfA was originally identified as a putative regulator of the activity of D. discoideum retrotransposon TRE5-A, it also was found to be essential for aggregation of D. discoideum cells. The identified DNA-binding activity was absent in mutant cells depleted of CbfA, and CbfA could be precipitated using an acaA promoter fragment. We propose that CbfA binds to the acaA promoter to provide a basal transcription activity that is required for induction of ACA expression after the onset of D. discoideum development.