Gene cloning, sequence, expression and in situ localization of 80 kDa diacylglycerol kinase specific to oligodendrocyte of rat brain

Goto, Kaoru; Watanabe, Masahiko; Kondo, Hisatake; Yuasa, H; Sakane, Fumio; Kanoh, Hideo

doi:10.1016/0169-328x(92)90196-i

Cited by 117 publications

(113 citation statements)

References 36 publications

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“…However, it is not known which DGK isozyme so far cloned accounts for the DGK activity variably distributed within the cells. In the present work we focussed on DGKc~, which is predominantly expressed in thymus and peripheral T-cells in addition to oligodendroglial cells [3][4][5].…”

Section: Introduction 2 Materials and Methodsmentioning

confidence: 99%

“…Rabbit anti-DGK(x antibody was raised against the N-terminal portion (amino acids 2 357) of rat DGKc~ and affinity-purified through an antigen column [5]. This antibody was previously shown to be specific to rat [5] and human [7] DGKc~.…”

Section: Materandismentioning

confidence: 99%

“…This antibody was previously shown to be specific to rat [5] and human [7] DGKc~. Anti-rat protein kinase Ca and anti-rat TcR antibodies were obtained from Gibco BRL and Serotech, Oxford, UK, respectively.…”

Section: Materandismentioning

confidence: 99%

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Translocation of diacylglycerol kinase α to the nuclear matrix of rat thymocytes and peripheral T‐lymphocytes

et al. 1996

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The cytosolic ~-diacylglycerol kinase (DGK) was translocated to and tightly associated with the nuclear matrix when rat thymocytes and peripheral T-lymphocytes were stimulated with concanavalin A or anti-T-cell receptor antibody. This translocation occurred rather slowly and was completed in 3-4 h after cell stimulation. We also detected significant accumulation of nuclear phosphatidic acid interpreted as being formed by the translocated enzyme. The enzyme translocation is not directly linked to pbospboinositide turnover and protein pbospborylation, since pborbol myristate acetate and calcium ionopbore did not affect the cellular DGK(x and since we detected no covalent modification of the enzyme molecule. Although the mechanisms underlying the enzyme translocation remain unknown, our results indicate that DGKcx participates in nuclear pbospbollpid metabolism occurring at the intermediate stage of lymphocyte activation.Key words." Diacylglycerol kinase, c~-isozyme; T-lymphocyte, rat; Translocation; Nuclear matrix elucidated by the finding that DGKa could account for the most of the DGK activity, which was rapidly increased after IL-2 stimulation of T-cell lines [18]. However, the role of DGKc~ in phosphoinositide turnover and in other biochemical events preceding the expression of IL-2 receptors remains unknown. For example, exogenous short-chain DG repeatedly administered to human T-lymphocytes was phosphorylated by cellular DGK only to a negligible extent [19]. Van der Bend et al. [20] proposed a topological sequestration of cellular DGK molecules based on the finding that DG artificially generated by treating Jurkat and other cells with bacterial phospholipase C could not serve as substrate for the cellular DGK. These observations indicate that the metabolic role of DGKa abundantly contained in T-lymphocytes is still largely unknown and that the cellular DGKs are operating under strict control mechanisms. Here we describe a targeting of DGKc~ to the nuclear matrix upon T-cell activation and suggest the participation of this DGK isozyme in the nuclear phospholipid metabolism occurring in stimulated lymphocytes.

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Section: Introduction 2 Materials and Methodsmentioning

confidence: 99%

Section: Materandismentioning

confidence: 99%

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Translocation of diacylglycerol kinase α to the nuclear matrix of rat thymocytes and peripheral T‐lymphocytes

et al. 1996

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“…The deduced 730-aa sequence (GenBank no. O88673) corresponds to a protein with a calculated M r of 82.7 kDa, with 92% sequence similarity to the 80-kDa rat DGK␣ (11), and 81% identity to the human and porcine counterpart (12,13). Like all type-I DGKs, this protein contains two EF hand motifs and two CRM, zinc finger-like sequences upstream of the C-terminal half, which shows considerable similarity with the C-terminal portion of all known DGK isoforms.…”

Section: Cloning and Expression Of Murine Dgk␣mentioning

confidence: 99%

“…DGK␣ protein expression was then analyzed in Western blot of extracts from murine tissues using a polyclonal Ab raised against DGK␣ of rat origin (11). The Ab recognized mouse DGK␣ expressed in COS cells (Fig.…”

Section: Cloning and Expression Of Murine Dgk␣mentioning

confidence: 99%

T Cell Activation In Vivo Targets Diacylglycerol Kinase α to the Membrane: A Novel Mechanism for Ras Attenuation

Sanjuán

Pradet‐Balade

Jones

et al. 2003

The Journal of Immunology

110

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Diacylglycerol kinase (DGK) phosphorylates diacylglycerol to produce phosphatidic acid, leading to decreased and increased levels, respectively, of these two lipid messengers that play a central role in T cell activation. Nine DGK isoforms, grouped into five subtypes, are found in higher organisms; all contain a conserved C-terminal domain and at least two cysteine-rich motifs of unknown function. In this study, we have researched in vivo the regulation of DGKα, using a transgenic mouse model in which injection of an antigenic peptide activates the majority of peripheral T cells. We demonstrate that DGKα, highly expressed in resting T lymphocytes, is subject to complex control at the mRNA and protein levels during in vivo T cell activation. Subcellular fractionation of T lymphocytes shortly after in vivo engagement of the TCR shows rapid translocation of cytosolic DGKα to the membrane fraction. At early time points, DGKα translocation to the membrane correlates with rapid translocation of Ras guanyl nucleotide-releasing protein (RasGRP), a nucleotide exchange activator for Ras that associates to the membrane through a diacylglycerol-binding domain. To demonstrate a causal relationship between DGKα activity and RasGRP relocation to the membrane, we determined RasGRP translocation kinetics in a T cell line transiently transfected with constitutive active and dominant-negative DGKα mutants. We show that membrane localization of DGKα is associated with a negative regulatory signal for Ras activation by reversing RasGRP translocation. This study is the first demonstration of in vivo regulation of DGKα, and provides new insight into the functional role of a member of this family of lipid kinases in the regulation of the immune response.

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Immunolocalization of Drosophila eye‐specific diacylgylcerol kinase, rdgA , which is essential for the maintenance of the photoreceptor

et al. 1997

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The Drosophila retinal degeneration A (rdgA) mutant has photoreceptor cells that degenerate within a week after eclosion. The degeneration starts with the disruption of the subrhabdomeric cisternae (SRC), which are the organelles essential for the transport of phospholipids to the photoreceptive membranes. Our previous biochemical and molecular studies suggested that the rdgA gene encodes an eye‐specific diacylglycerol kinase (DGK). In this study, we show that retinal degeneration is prevented by the introduction of the eye‐DGK gene in the rdgA mutant genome, suggesting that the DGK activity is crucial for the maintenance of the photoreceptor. Furthermore, by immunohistochemical analysis, we have demonstrated that the rdgA protein is predominantly associated with the SRC, suggesting that the conversion from diacylglycerol (DG) to phosphatidic acid (PA) most actively occurs in SRC. The analysis of the eyes of mutants homozygous for rdgA and eye‐protein kinase C mutations indicates that retinal degeneration is caused by the deficiency of PA rather than excessive accumulation of DG. From these data, we conclude that the production of PA in the SRC membranes is essential for the maintenance of the photoreceptor. © 1997 John Wiley & Sons, Inc. J Neurobiol 32: 695–706, 1997

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Gene cloning, sequence, expression and in situ localization of 80 kDa diacylglycerol kinase specific to oligodendrocyte of rat brain

Cited by 117 publications

References 36 publications

Translocation of diacylglycerol kinase α to the nuclear matrix of rat thymocytes and peripheral T‐lymphocytes

Translocation of diacylglycerol kinase α to the nuclear matrix of rat thymocytes and peripheral T‐lymphocytes

T Cell Activation In Vivo Targets Diacylglycerol Kinase α to the Membrane: A Novel Mechanism for Ras Attenuation

Immunolocalization of Drosophila eye‐specific diacylgylcerol kinase, rdgA , which is essential for the maintenance of the photoreceptor

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