Signal transduction for apoptosis induction using apoptosis-inducing immunosuppressant

使用细胞凋亡诱导免疫抑制剂诱导细胞凋亡的信号转导

• 批准号: 09680588
• 负责人: KOZUTSUMI Yasunori
• 金额: $ 1.86万
• 依托单位: KYOTO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09680588/
• 关键词: apoptosis; immunosuppressant; glycolipid; sphingolipid; kinase; signal transduction; ISP-1; yeast; スフィンゴシン; CPP32; Bcl-2

In our previous study, the sphingosine-like immunosuppressant, ISP-1, was found to suppress proliferation of an interleukin-2-depentent cytotoxic T cell line, CTLL-2, through the inhibition of serine palmitoyltransferase, which catalyzes the committed step of sphingolipid biosynthesis. The growth inhibition induced by ISP-1 was antagonized by the addition of sphingosine. Instead of sphingosine, phytosphingsine is synthesized in the Saccharomyces cerevisiae by the similar pathway of mammalian cells, In this study, the inhibition of proliferation by ISP-1 was observed with Saccharomyces cerevisiae, and was abolished by addition of phytosphingsine. In addition, overreplication of DNA was detected in the ISP-1 treated yeast cells by flow cytometry. And we have identified a Saccharomyces cerevisiae gene, SLI2, that resisted the growth inhibition of ISP-1. The protein kinase catalytic domain is located at the carboxy-terminal portion of Sli2p. Kinase-dead mutants of SLI2 did not give any resistance to ISP-1, leading us to predict that kinase activity of Sli2p should be essential for its function in resisting ISP-1. Sli2p overexpression had no effect on serine palmitoyltransferase activity and sphingolipids metabolism. These results indicated that Sli2p may play an important role in rescuing yeast from decrease of sphingolipids induced by ISP-1.

在我们先前的研究中，发现类似鞘氨酸样免疫抑制剂ISP-1，可以通过抑制丝氨酸棕榈酰转移酶的抑制，抑制白介素-2-二型细胞毒性T细胞CTLL-2的增殖，从而催化丝网膜含量的步骤生物合成。 ISP-1诱导的生长抑制是通过添加鞘氨醇拮抗的。代替鞘氨氨酸，植物磷氨酸是通过哺乳动物细胞的相似途径在酿酒酵母中合成的，在这项研究中，用酿酒酵母观察到ISP-1抑制增殖，并通过添加植物磷酸化。另外，通过流式细胞仪在ISP-1处理的酵母细胞中检测到DNA的过度复制。我们已经确定了酿酒酵母基因SLI2，该基因抗拒ISP-1的生长抑制作用。蛋白激酶催化结构域位于SLI2P的羧基末端部分。 SLI2的激酶死亡突变体对ISP-1没有任何抗性，这使我们预测SLI2P的激酶活性对于抵抗ISP-1的功能至关重要。 SLI2P过表达对丝氨酸棕榈酰转移酶活性和鞘脂代谢没有影响。这些结果表明，SLI2P可能在挽救ISP-1诱导的鞘脂降低酵母中起重要作用。

项目成果

K., Terayama et al.: "Cloning and functional expression of a novelglucuronyltransferase involved in the biosynthesis of the carbohydrate epitope, HNK-1" Proc. Natl. Acad. Sci. U.S.A.94(12). 6093-6098 (1997)

K.、Terayama 等人：“参与碳水化合物表位 HNK-1 生物合成的新型葡萄糖醛酸转移酶的克隆和功能表达”Proc.

H., Morio et al.: "Renal expression of serum-type mannan-binding protein in rat." Eur.J.Biochem.243(3). 770-774 (1997)

H.，Morio 等人：“大鼠血清型甘露聚糖结合蛋白的肾脏表达。”

山地俊之他2名: "免疫抑制剤ISP-1の作用機構" 蛋白質核酸酵素. 43(16). 2503-2509 (1998)

Toshiyuki Yamaji 等 2 人：“免疫抑制剂 ISP-1 的作用机制”蛋白质核酸酶 43(16) (1998)。

K.Terayama, S.Oka, Y.Miki, A.Nakamura, Y.Kozutsumi, K.Takio and T.Kawasaki: "Cloning and functional expression of a novel glucuronyltransferase involved in the biosynthesis of the carbohydrate epitope, HNK-1" Proc.Natl.Acad.Sci.U.S.A.94(12). 6093-6098 (19

K.Terayama、S.Oka、Y.Miki、A.Nakamura、Y.Kozutsumi、K.Takio 和 T.Kawasaki：“参与碳水化合物表位 HNK-1 生物合成的新型葡萄糖醛酸转移酶的克隆和功能表达”

H.Sakuma, A.Ikeda, s.Oka, Y.Kozutsumi, J-P.Zanetta, and T.Kawasaki: "Molecular Cloning and Functional Expression of a cDNA Encoding a New member of Mixed Lineage protein Kinase from Human Brain" J.Biol.Chem.272(45). 28622-28629 (1997)

H.Sakuma、A.Ikeda、s.Oka、Y.Kozutsumi、J-P.Zanetta 和 T.Kawasaki：“编码人脑混合谱系蛋白激酶新成员的 cDNA 的分子克隆和功能表达”J.Biol