The rbp9 gene of Drosophila melanogaster has been molecularly characterized and shown to be expressed solely in the CNS, where it encodes proteins with three RNA recognition motifs (RRMs). Sequencing of genomic and cDNA clones of rbp9 revealed a complex gene with three alternative promoters as well as alternative patterns of splicing. The deduced amino acid sequence of the RBP9 proteins is highly similar to those of three other nervous system-specific genes, human HuC and HuD and Drosophila elav, which also encode proteins with three RRMs. Developmental Northern analysis revealed that rbp9 is expressed from the late third instar larva through adult stages. The RBP9 protein was detected specifically in nuclei of the nervous system after morphogenesis of the adult CNS in the mid-pupal stage. Thus, the RBP9 protein does not appear until substantially later than rbp9 transcripts are detected. The adult nervous system nuclear-limited expression pattern, the presence of RRMs, and the high similarity to a group of nervous system-specific proteins in flies and humans suggest that rbp9 belongs to a nervous system-specific RRM protein gene subfamily that may participate in the processing of RNAs involved in the development of the CNS.