Vertebrate tissue inhibitors of metalloproteinases (TIMPs) regulate extracellular matrix metalloproteinases and are thus involved in a wide variety of developmental and physiological processes. By identifying cDNAs of a transcript detected within an intron of the Drosophila synapsin gene we have cloned the Drosophila TIMP gene (Timp), which represents the first invertebrate member of the TIMP gene family. Sequence analysis revealed an open reading frame of 210 amino acids with 35% identity to human TIMPs and a conserved exon-intron structure. Analysis of sequence data from the Sanger Centre demonstrated that the human TIMP3 gene is encoded within intron V of the human synapsin-III gene, indicating that the nested organization of TIMP and synapsin genes may be a general feature conserved in evolution. We therefore speculate that the human TIMP4 gene will be located in intron V of the human synapsin-II gene on chromosome 3p25, and we present preliminary evidence that a human synapsin-IV gene is located near the TIMP2 gene on chromosome 17q23-q25.