The genomic and cDNA fragments of Drosophila melanogaster, homologous to human c-raf-1, were cloned. The nucleotide sequence predicted the primary structure of a polypeptide of 666 amino acid residues with a highly conserved Ser-Thr kinase domain on its carboxy terminal half. Draf-1 was mapped to the 2F region of the X chromosome. Two newly induced recessive lethals belonging to a complementation group in this region were identified to be defective in Draf-1 by P element-mediated rescue experiments. The mutants die at larval/pupal stages. The mutant larvae are apparently normal, but they harbor serious defects in the organs containing proliferating cells of both somatic and germ line origins. Maternal effects on embryogenesis indicated that Draf-1 is also required in early larval development.