The synthesis and utilization of CDP-diacylglycerol in mammalian cells was demonstrated over 35 years ago when initial studies were carried out. However, CDP-diacylglycerol synthases and the genes encoding these enzymes have been studied in the greatest detail in Escherichia coli and Saccharomyces cerevisiae. The involvement of CDP-diacylglycerol in regulation of phospholipid metabolism has recently been demonstrated in Saccharomyces cerevisiae, and evidence now exists from studies in Drosophila that this liponucleotide may be important in regulation of lipid-dependent signal transduction processes. The vast amount of biochemical and genetic information on the synthases from microorganisms has led to the cloning of genes that encode CDP-diacylglycerol synthases from somatic cells. The combination of information on these synthases from all organisms will lead to a clearer understanding of the role CDP-diacylglycerol plays in cellular processes.