Muskelin was identified in vertebrates as a novel, intracellular, kelch repeat protein that is needed in cell-spreading responses to the matrix adhesion molecule, thrombospondin-1. The identification and characterization of an orthologue of muskelin in Drosophila melanogaster is now reported. The Drosophila muskelin gene, located on chromosome 2R, is encoded in ten exons. Drosophila muskelin is expressed in embryos, larvae and adult flies. The protein has 45% sequence identity to vertebrate muskelins, with highest sequence identity in an amino-terminal domain and the six kelch repeats that form a beta-propeller structure. Multiple sequence alignment of human, mouse, rat and Drosophila muskelins and protein database searches revealed a novel highly conserved motif within the amino-terminal domain, lissencephaly homology motif (LisH) and C-terminal to LisH motifs in the central region of the molecule, and several conserved consensus motifs for phosphorylation by protein kinase C and casein kinase II. These findings provide new information on the modular structure of muskelin and indicate potential for conserved mechanisms of function.