N,N-Dimethylsphingosine br Funding br Conflict of interests

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Introduction DAG is generated by the hydrolysis of phosphatidylinositol-4,5-bisphosphate (PtdIns(4,5)P2) by PtdIns-specific phospholipase C (PLC) enzymes [1]. Remaining in the membrane, it binds proteins with cysteine-rich, C1 domains, and activates several of these proteins including protein kinase C (PKC) isoforms [2] and Ras guanyl nucleotide-releasing proteins (RasGRPs) [3]. In other cases, DAG recruits but does not activate proteins such as protein kinase D, the Munc13 proteins, and the chimaerins [3]. Additionally, DAG appears to activate some transient receptor potential N,N-Dimethylsphingosine that do not harbor C1 domains [4]. Its effects on numerous and diverse targets underscores the importance of DAG signaling and indicates that DAG modulates a broad array of biological events. It is critical then that intracellular DAG levels be tightly regulated and it is now widely agreed that under most circumstances, conversion of DAG to PA by the DGKs is the major route to terminate DAG signaling. But PA, itself, has a broad array of signaling properties that are very distinct from those of DAG. Their ability to generate PA suggests that DGKs might also influence biological events by generating this lipid. In fact, there are now several examples indicating that DGKs modulate signaling events not only by metabolizing DAG to terminate its effects, but also by producing PA. DGKs have been identified in unicellular organisms such as bacteria, but these DGKs are structurally different from those identified in higher eukaryotes such as Drosophila melanogaster[5], [6], [7], Arabidopsis thaliana[8], [9], Caenorhabditis elegans[10], [11], and mammals [12]. For example, bacterial DGK, unlike higher eukaryote DGKs, is a small, integral membrane protein that phosphorylates other lipids in addition to DAG [13]. And a recently identified yeast DGK is similar to cytidyltransferases and uses CTP as a phosphate donor rather than the ATP used by DGKs in higher eukaryotes [14], [15]. Finally, a multisubstrate lipid kinase (MuLK) that also phosphorylates DAG was recently identified [16], [17]. Unlike mammalian DGKs, MuLK phosphorylates non-DAG lipids and its structure is not very homologous to that of DGKs. This review will focus on DGKs in higher eukaryotes, with a specific focus on their role in generating PA that influences biological events.
Common structural features of the mammalian DGK enzymes DGK activity was first identified about forty years ago [18], and an 80 kDa DGK enzyme was purified to homogeneity from pig brain cytosol in 1983 [19]. Based on the partial sequence from the 80 kDa enzyme, Sakane et al. isolated a full-length, porcine DGK cDNA in 1990 [20]. But based on antibody studies, additional DGK enzymes appeared to exist [21], prompting an exhaustive search for other DGK isoforms. Nine additional isoforms have since been identified (Fig. 1), making this a rather large family of enzymes. Their diversity is even broader because DGKs β, γ, δ, ζ, ι, and η are alternatively spliced [22]. All mammalian DGKs have two common structural elements: a catalytic domain and at least two C1 domains. The functions and structural properties of these domains have been described in detail in recent reviews [22], [23], so only the important features of these motifs are highlighted below.
DGKs influence specific biological events depending on their binding partners Each DGK isotype is expressed in numerous tissues, and usually multiple DGK isotypes are expressed in the same tissue and even within the same cell [50]. For example we detected all known DGK isoforms in mouse brain extracts [51] and have found expression of at least six DGK isoforms in mouse embryo fibroblasts (J.C. and M.K.T. unpublished observations). When multiple DGK isoforms are expressed in a cell type, they are usually from different subfamilies, suggesting that the subfamilies have distinct functional roles. Thus, specific pools of DAG could be uniquely regulated by directing DGK isoforms to appropriate cellular compartments in order to metabolize the DAG. Conversely, DGK isotypes could uniquely generate PA depending on their intracellular localization and modes of regulation. Consistent with this model, evidence indicates that DGKs achieve functional specificity by accessing specific pools of DAG and binding to a unique subset of DAG- or PA-activated proteins in order to regulate their activity (Fig. 2). This concept agrees with an emerging body of evidence indicating that specificity in signal transduction is often achieved by gathering together signaling proteins in common pathways along with their regulators [52].