Cloning of a novel human diacylglycerol kinase (DGKθ) containing three cysteine-rich domains, a proline-rich region, and a pleckstrin homology domain with an overlapping Ras-associating domain

Abstract

Diacylglycerol kinase (DGK) attenuates levels of second messenger diacylglycerol in cells and produces another (putative) messenger, phosphatidic acid. We have previously purified a 110-kDa DGK from rat brain (Kato, M., and Takenawa, T. (1990) J. Biol. Chem. 265, 794-800). Here we report the cDNA cloning from human brain and retina cDNA libraries. The cDNA encodes a novel DGK isotype, termed DGKΘ, of 941 amino acids with an apparent molecular mass of 110 kDa. DGKΘ contains a C-terminal putative catalytic domain, which is present in all eukaryotic DGKs. In contrast to other DGK isotypes, DGKΘ contains three cysteine-rich domains instead of two. The third cysteine.rich domain is most homologous to the second one in other DGK isotypes. This particular sequence homology extends C-terminally beyond the typical cysteine/histidine core structure and is DGK-specific. DGKΘ furthermore contains various domains for protein-protein interaction, such as a proline- and glycine-rich domain with a putative SH3 domain- binding site and a pleckstrin homology domain with an overlapping Ras- associating domain. DGKΘ is expressed in the brain and, to a lesser extent, in the small intestine, duodenum, and liver. In situ hybridization of DGKΘ mRNA in adult rat brain reveals high expression in the cerebellar cortex and hippocampus. DGKΘ activity in COS cell lysates is optimal toward diacylglycerols containing an unsaturated fatty acid at the sn-2 position.