Role of PIP2 and PIP3 binding proteins on cell growth signals

PIP2 和 PIP3 结合蛋白对细胞生长信号的作用

• 批准号: 06404022
• 负责人: TAKENAWA Tadaomi
• 金额: $ 18.69万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-06404022/
• 关键词: alpha-actinin; PIP_2; PH domain; PIP_3; ホスホリパーゼC; 細胞骨格

1. Histons as novel PIP_2 binding proteinsWe found that PIP_2 bound to histon H_1 and H_3 and determined the binding sites. PIP_2 bound to the C-terminal 103 aminoacids. The amounts of PIP_2 bound to histon H_1 decreased when histon H_1 was phosphorylated by PKC.The addition of PIP_2 to histon H_1 suppressed the inhibitory effect of histon H_1 on basic transcriptional activity by RNA polymerase.2. PIP_2 phosphatase downstream of tyrosine kinaseWe purified 150kDa protein bound to Ash/Grb2 from bovine brain, and isolated its cDNA.P150 was found to have homology with synaptojanin. When P150 was expressed in Cos cells, actin stress fibers was disrupted and the cells showed multi-nuclear in shape. We purified P150 from bovine brain and characterized it. PIP_2 phosphatase activity was not inhibited by actin regulatory proteins such as cofilin, profilin and alpha-actinin, While PLCdelta_1 was inhibited. These results suggest that P150 hydrolyzes PIP_2 bound to actin regulatory proteins, resulting in reorganization of actin fibers.

1.组蛋白作为新的PIP_2结合蛋白我们发现PIP_2与组蛋白希斯顿H_1和H_3结合，并确定了结合位点。PIP_2与C-末端103个氨基酸结合。当希斯顿H_1被PKC磷酸化后，PIP_2与希斯顿H_1的结合量减少，PIP_2与希斯顿H_1的结合抑制了希斯顿H_1对RNA聚合酶基础转录活性的抑制作用. PIP_2酪氨酸激酶下游磷酸酶我们从牛脑中纯化了与Ash/Grb 2结合的150 kDa蛋白，并分离了其cDNA，发现P150与synaptojanin具有同源性。当P150在Cos细胞中表达时，肌动蛋白应力纤维被破坏，细胞呈多核状。我们从牛脑中分离纯化了P150，并对其进行了鉴定，发现PIP_2磷酸酶活性不受肌动蛋白调节蛋白如cofilin、profilin和α-actinin的抑制，而PLC δ_1则受到抑制。这些结果表明，P150水解与肌动蛋白调节蛋白结合的PIP_2，导致肌动蛋白纤维的重组。

项目成果

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K.Fukami、T.Endo、M.Imamura 和 T.Takenawa：“α-肌动蛋白和纽蛋白是参与酪氨酸激酶信号传导的 PIP2 结合蛋白”J.Biol.Chem.269 1518-1522 (1994)。

H.Tsubokawa,K.Oguro,H.P.C.Robinson,T.Matuzawa,S.G.Rhee,T.Takenawa and N.Kawai: "Inositol 1,3,4,5-tetrakisphosphate as a mediator of neuronal death in ischemic hippocampus" Neuroscience. 59. 291-297 (1994)

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A.Ando、K.Yonezawa、I.Gout、T.NaKta、H.Ueda、K.Hara、Y.Kitamura、Y.Noda、T.Takenawa、N.Hirokawa、M.Waterfield 和 M.Kasuga：“A

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