title: Cloning of a novel human diacylglycerol kinase (DGKtheta) 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 DGKtheta, of 941 amino acids with an apparent molecular mass of 110 kDa. DGKtheta contains a C-terminal putative catalytic domain, which is present in all eukaryotic DGKs. In contrast to other DGK isotypes, DGKtheta 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. DGKtheta 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. DGKtheta is expressed in the brain and, to a lesser extent, in the small intestine, duodenum, and liver. In situ hybridization of DGKtheta mRNA in adult rat brain reveals high expression in the cerebellar cortex and hippocampus. DGKtheta activity in COS cell lysates is optimal toward diacylglycerols containing an unsaturated fatty acid at the sn-2 position.
authors.affiliation: Division of Cellular Biochemistry, Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam, The Netherlands.
authors.forename: B
authors.initials: B
authors.lastname: Houssa
authors.forename: D
authors.initials: D
authors.lastname: Schaap
authors.forename: J
authors.initials: J
authors.lastname: van der Wal
authors.forename: K
authors.initials: K
authors.lastname: Goto
authors.forename: H
authors.initials: H
authors.lastname: Kondo
authors.forename: A
authors.initials: A
authors.lastname: Yamakawa
authors.forename: M
authors.initials: M
authors.lastname: Shibata
authors.forename: T
authors.initials: T
authors.lastname: Takenawa
authors.forename: W J
authors.initials: WJ
authors.lastname: van Blitterswijk
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D001798:Blood Proteins / Q000737:chemistry*; D001921:Brain / Q000201:enzymology*; D019556:COS Cells; D003001:Cloning, Molecular; D003545:Cysteine; D019852:Diacylglycerol Kinase; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D009928:Organ Specificity; D010446:Peptide Fragments / Q000737:chemistry; D010750:Phosphoproteins; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000096:biosynthesis* / Q000737:chemistry* / Q000378:metabolism; D011392:Proline; D051381:Rats; D011994:Recombinant Proteins / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity; D014162:Transfection; D018631:ras Proteins / Q000737:chemistry*
chemical_list: D001798:Blood Proteins; D015345:Oligonucleotide Probes; D010446:Peptide Fragments; D010750:Phosphoproteins; D011994:Recombinant Proteins; C039077:platelet protein P47; D011392:Proline; D017853:Phosphotransferases (Alcohol Group Acceptor); D019852:Diacylglycerol Kinase; D018631:ras Proteins; D003545:Cysteine