De La Roche Marc A, Smith Janet L, Rico Maribel, Carrasco Silvia, Merida Isabel, Licate Lucila, Côté Graham P, Egelhoff Thomas T
Department of Biochemistry, Queen's University, Kingston, Ontario, Canada K7L 3N6.
Biochem J. 2002 Dec 15;368(Pt 3):809-15. doi: 10.1042/BJ20021027.
Diacylglycerol kinases (DGKs) phosphorylate the neutral lipid diacylglycerol (DG) to produce phosphatidic acid (PA). In mammalian systems DGKs are a complex family of at least nine isoforms that are thought to participate in down-regulation of DG-based signalling pathways and perhaps activation of PA-stimulated signalling events. We report here that the simple protozoan amoeba Dictyostelium discoideum appears to contain a single gene encoding a DGK enzyme. This gene, dgkA, encodes a deduced protein that contains three C1-type cysteine-rich repeats, a DGK catalytic domain most closely related to the theta subtype of mammalian DGKs and a C-terminal segment containing a proline/glutamine-rich region and a large aspargine-repeat region. This gene corresponds to a previously reported myosin II heavy chain kinase designated myosin heavy chain-protein kinase C (MHC-PKC), but our analysis clearly demonstrates that this protein does not, as suggested by earlier data, contain a protein kinase catalytic domain. A FLAG-tagged version of DgkA expressed in Dictyostelium displayed robust DGK activity. Earlier studies indicating that disruption of this locus alters myosin II assembly levels in Dictyostelium raise the intriguing possibility that DG and/or PA metabolism may play a role in controlling myosin II assembly in this system.
二酰基甘油激酶(DGKs)将中性脂质二酰基甘油(DG)磷酸化以产生磷脂酸(PA)。在哺乳动物系统中,DGKs是一个复杂的家族,至少有九种亚型,被认为参与基于DG的信号通路的下调,也可能参与PA刺激的信号事件的激活。我们在此报告,简单的原生动物变形虫盘基网柄菌似乎含有一个编码DGK酶的单一基因。这个基因,即dgkA,编码一种推导的蛋白质,它包含三个C1型富含半胱氨酸的重复序列、一个与哺乳动物DGKs的θ亚型最密切相关的DGK催化结构域以及一个包含富含脯氨酸/谷氨酰胺区域和一个大的天冬酰胺重复区域的C末端片段。这个基因对应于先前报道的肌球蛋白II重链激酶,称为肌球蛋白重链-蛋白激酶C(MHC-PKC),但我们的分析清楚地表明,正如早期数据所暗示的那样,该蛋白质并不包含蛋白激酶催化结构域。在盘基网柄菌中表达的带有FLAG标签的DgkA版本表现出强大的DGK活性。早期研究表明,该基因座的破坏会改变盘基网柄菌中肌球蛋白II的组装水平,这引发了一个有趣的可能性,即DG和/或PA代谢可能在该系统中控制肌球蛋白II的组装中发挥作用。