Distinct expression and localization of the type II diacylglycerol kinase isozymes δ, η and κ in the mouse reproductive organs

Shionoya, Takao; Usuki, Takako; Komenoi, Suguru; Isozaki, Tetsuro; Sakai, Hiromichi; Sakane, Fumio

doi:10.1186/s12861-015-0055-z

Cited by 20 publications

(12 citation statements)

References 39 publications

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“…DGK is abundantly expressed in the testis (14,30). A genome-wide association study indicated a potential relationship between DGK and hypospadias (31).…”

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confidence: 99%

The Pleckstrin Homology Domain of Diacylglycerol Kinase η Strongly and Selectively Binds to Phosphatidylinositol 4,5-Bisphosphate

Kume

Kawase

Komenoi

et al. 2016

Journal of Biological Chemistry

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Type II diacylglycerol kinase (DGK) isozymes (␦, , and ) have a pleckstrin homology domain (PH) at their N termini. Here, we investigated the lipid binding properties of the PHs of type II DGK isozymes using protein-lipid overlay and liposome binding assays. The PH of DGK showed the most pronounced binding activity to phosphatidylinositol (PI) 4,5-bisphosphate (PI(4,5)P 2 ) among the various glycero-and sphingolipids including PI 3,4,5-trisphosphate, PI 3,4-bisphosphate, PI 3-phosphate, PI 4-phosphate, and PI 5-phosphate. Moreover, the PI(4,5)P 2 binding activity of the DGK-PH was significantly stronger than that of other type II DGK isozymes. Notably, compared with the PH of phospholipase C (PLC) ␦1, which is generally utilized as a cellular PI(4,5)P 2 -probe, the DGK-PH is equal to or superior than the PLC␦1-PH in terms of affinity and selectivity for PI(4,5)P 2 . Furthermore, in COS-7 cells, GFP-fused wild-type DGK1 and its PH partly translocated from the cytoplasm to the plasma membrane where the PLC␦1-PH was co-localized in response to hyperosmotic stress in an inositol 5-phosphatase-sensitive manner, whereas a PH deletion mutant did not. Moreover, K74A and R85A mutants of DGK-PH, which lack the conserved basic amino acids thought to ligate PI(4,5)P 2 , were indeed unable to bind to PI(4,5)P 2 and co-localize with the PLC␦1-PH even in osmotically shocked cells. Overexpression of wild-type DGK1 enhanced EGF-dependent phosphorylation of ERK, whereas either K74A or R85A mutant did not. Taken together, these results indicate that the DGK-PH preferentially interacts with PI(4,5)P 2 and has crucial roles in regulating the subcellular localization and physiological function of DGK. Moreover, the DGK-PH could serve as an excellent cellular sensor for PI(4,5)P 2 . Diacylglycerol kinase (DGK)2 phosphorylates diacylglycerol to produce phosphatidic acid (1-5). Diacylglycerol (6 -8) and phosphatidic acid (PA) (9, 10) are well recognized as lipid second messengers. DGK appears to participate in various physiological events by modulating the balance between the two bioactive lipids diacylglycerol and phosphatidic acid (1-5). Ten mammalian DGK isozymes (␣, ␤, ␥, ␦, ⑀, , , , , and ) containing two or three characteristic zinc finger-like C1 domains and a common catalytic region are divided into five groups (types I-V) according to their structural features (1-5).Type II DGKs (11) consist of DGK␦ (12), - (13), and -(14). Moreover, alternative splicing products of DGK␦ (␦1 and ␦2) (15) and -(1 and 2) (16) have been identified. Type II DGKs have a pleckstrin homology domain (PH) in common at their N termini and a catalytic domain that is divided into two subdomains (catalytic subdomains a and b). DGKs ␦1, ␦2, and 2, but not DGKs 1 and , contain a sterile ␣-motif domain at their C termini. It has been demonstrated that DGKs ␦1, ␦2, and 2 form oligomers through interactions between their sterile ␣-motif domains and that this oligomer formation regulates the activities and subcellular localization of these DGK isoforms (15)(16)(1...

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“…DGK is abundantly expressed in the testis (14,30). A genome-wide association study indicated a potential relationship between DGK and hypospadias (31).…”

mentioning

confidence: 99%

The Pleckstrin Homology Domain of Diacylglycerol Kinase η Strongly and Selectively Binds to Phosphatidylinositol 4,5-Bisphosphate

Kume

Kawase

Komenoi

et al. 2016

Journal of Biological Chemistry

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“…Although the DGKη3 mRNA was strongly expressed in the testis, its protein band has not been detected in our previous report [ 25 ]. It is possible that the DGKη3 protein is unstable and quickly degraded.…”

Section: Resultsmentioning

confidence: 64%

“…We previously found a new alternative splicing product of DGKη gene, DGKη3, which lacks exon 26 encoding 31 amino acid residues [ 25 ] ( Fig 1 ). However, it is still unknown whether the DGKη3 mRNA is a derivative of the DGKη1 mRNA or the DGKη2 mRNA.…”

Section: Resultsmentioning

confidence: 99%

“…Several reports have revealed that DGKη is abundantly expressed in the reproductive organs, testis and ovary [ 8 – 12 ]. Therefore, we examined the expression and spatial distribution of the DGKη1 and η2 proteins and mRNAs in the mouse reproductive organs [ 25 ]. The results indicate distinct expression patterns, which were obviously different from each other.…”

Section: Introductionmentioning

confidence: 99%

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Cloning and Characterization of Novel Testis-Specific Diacylglycerol Kinase η Splice Variants 3 and 4

et al. 2016

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Diacylglycerol kinase (DGK) phosphorylates DG to generate phosphatidic acid. Recently, we found that a new alternative splicing product of the DGKη gene, DGKη3, which lacks exon 26 encoding 31 amino acid residues, was expressed only in the secondary spermatocytes and round spermatids of the testis. In this study, we cloned the full length DGKη3 gene and confirmed the endogenous expression of its protein product. During the cloning procedure, we found a new testis-specific alternative splicing product of the DGKη gene, DGKη4, which lacks half of the catalytic domain. We examined the DGK activity and subcellular localization of DGKη3 and η4. DGKη3 had almost the same activity as DGKη1, whereas the activity of DGKη4 was not detectable. In resting NEC8 cells (human testicular germ cell tumor cell line), DGKη1, η3 and η4 were broadly distributed in the cytoplasm. When osmotically shocked, DGKη1 and η4 were distributed in punctate vesicles in the cytoplasm. In contrast, DGKη3 was partly translocated to the plasma membrane and co-localized with the actin cytoskeleton. These results suggest that DGKη3 and η4 have properties different from those of DGKη1 and that they play roles in the testis in a different manner.

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“…1C). Conversely, the DGK2 expression level in the testis, in which the protein was strongly expressed in wild-type mice (Shionoya et al, 2015;Usuki et al, 2015), was almost the same as that of the control littermates. We verified that deleting DGK did not significantly affect protein expression of other DGK isozymes (, , , , , , ,  and ) in the brain (Suppl.…”

Section: Generation Of Brain-specific Dgk-deficient Micementioning

confidence: 81%

Behavioral and pharmacological phenotypes of brain-specific diacylglycerol kinase δ-knockout mice

et al. 2016

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Diacylglycerol kinase (DGK) is a lipid-metabolizing enzyme that phosphorylates diacylglycerol to produce phosphatidic acid. Previously, we reported that the δ isozyme of DGK was abundantly expressed in the mouse brain. However, the functions of DGKδ in the brain are still unclear. Because conventional DGKδ-knockout (KO) mice die within 24h after birth, we have generated brain-specific conditional DGKδ-KO mice to circumvent the lethality. In the novel object recognition test, the number of contacts in the DGKδ-KO mice to novel and familiar objects was greatly increased compared to the control mice, indicating that the DGKδ-KO mice showed irrational contacts with objects such as compulsive checking. In the marble burying test, which is used for analyzing obsessive-compulsive disorder (OCD)-like phenotypes, the DGKδ-KO mice buried more marbles than the control mice. Additionally, these phenotypes were significantly alleviated by the administration of an OCD remedy, fluoxetine. These results indicate that the DGKδ-KO mice showed OCD-like behaviors. Moreover, the number of long axon/neurites increased in both DGKδ-KO primary cortical neurons and DGKδ-knockdown neuroblastoma Neuro-2a cells compared to control cells. Conversely, overexpression of DGKδ decreased the number of long axon/neurites of Neuro-2a cells. Taken together, these results strongly suggest that a deficiency of DGKδ induces OCD-like behavior through enhancing axon/neurite outgrowth.

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Distinct expression and localization of the type II diacylglycerol kinase isozymes δ, η and κ in the mouse reproductive organs

Cited by 20 publications

References 39 publications

The Pleckstrin Homology Domain of Diacylglycerol Kinase η Strongly and Selectively Binds to Phosphatidylinositol 4,5-Bisphosphate

The Pleckstrin Homology Domain of Diacylglycerol Kinase η Strongly and Selectively Binds to Phosphatidylinositol 4,5-Bisphosphate

Cloning and Characterization of Novel Testis-Specific Diacylglycerol Kinase η Splice Variants 3 and 4

Behavioral and pharmacological phenotypes of brain-specific diacylglycerol kinase δ-knockout mice

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