Three human genes encoding proteins homologous to Drosophila Dishevelled protein were cloned and characterized. Amino acid similarity between the different Dishevelled proteins is concentrated in three highly conserved regions. Two of these regions do not exhibit significant sequence similarity with other known proteins; the third is similar to the discs-large homology region, which was first found in a Drosophila Discs-large tumor suppressor protein (also known as GLGF or PDZ domain). We produced antibodies against human Dishevelled-2 and demonstrated that it is a phosphoprotein and can be detected in all cell lines and human embryonic tissues examined. Indirect immunofluorescence indicates that it is found throughout the cytoplasm. Our results indicate that the human dishevelled genes constitute a multigene family and that Dishevelled proteins are highly conserved among metazoans.