Regulatgion of translation by protein phosphorylation.

通过蛋白质磷酸化调节翻译。

• 批准号: 12680623
• 负责人: SHIMA Hiroshi
• 金额: $ 2.69万
• 依托单位: HOKKAIDO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12680623/
• 关键词: S6; phosphorylation; kinase; phosphatase; PP1; PP2A; calcineurin; tautomycetin; S6キナーゼ; 欠損マウス; トウトマイシン; 細胞増殖; 癌化; 翻訳; がん化; 細胞周期; 免疫抑制

1. Regulation of translation by type 1 protein phosphatase (PP1 )PP1 is thought to be involved in dephosphorylation and negative regulation of S6 protein. Two kinds of PP1 regulatory proteins (NIPP-1 and 1-4) were identified and characterized. NIPP-1 was shown to bind 4 kinds of PP1 catalytic subunits with similar affinity. Expression of NIPP-1 mRNA in rat hepatomas was analyzed and It was shown that its level was positively correlated with the malignant phenotype.We have isolated a novel and the most potent PP1 inhibitory protein named as I-4. IC50 towards PP1 of is 0.2hM. The experiments based on deletion mutants showed that I-4 inhibits PP1 by covering catalytic site of PP1 catalytic subunit by binding with 3 domains.2. S6 kinase deficient mouseBased on the phenotype of S6 kinase deficient mouse, we reveled a novel isotope of S6K, S6K2. Development and analysis of S6K2 deficient mouse is currently, on going.3. Regulation of translation by type 2A protein phosphatase (PP2A)APP2A regulatory protein (I-2/PP2A) was isolated. The expression of I-2/PP2A mRNA was regulated up regulated at G1/S. Over-expression of I-2/PP2A suppressed the cell proliferation.4. Mechanism of immunosuppressive drugAlthough the target of immunosuppressive drug such as cyclosporinA and FK506 is identified as calcineurin, the assay method for calcineurin activity has not been established well. We established the efficient and accurate method for enzyme activity of calcineurin.5. Cross-talk between translation and major signal cascadesWe have identified novel negative regulator named as MKP-7 (for MAPK pathway) and PTP epsilon (for JAK-STAT) and cauterized the mode of action of these proteins.

1. 1型蛋白磷酸酶（PP 1）对翻译的调节被认为参与了S6蛋白的去磷酸化和负调节。对两种PP 1调节蛋白（NIPP-1和1-4）进行了鉴定和鉴定。NIPP-1与4种PP 1催化亚基的结合亲和力相似。本研究分析了NIPP-1 mRNA在大鼠肝癌中的表达，结果表明其表达水平与肝癌恶性表型呈正相关，我们分离到一种新的、最有效的PP 1抑制蛋白，命名为I-4。对PP 1的IC 50为0.2hM。基于缺失突变体的实验表明，I-4通过与3个结构域结合覆盖PP 1催化亚基的催化位点来抑制PP 1. S6激酶缺陷型小鼠根据S6激酶缺陷型小鼠的表型，我们发现了一种新的S6 K同位素S6 K2。S6 K2缺陷小鼠的开发和分析目前正在进行中. 2A型蛋白磷酸酶（PP 2A）对翻译的调节分离出APP 2A调节蛋白（I-2/PP 2A）。I-2/PP 2A mRNA在G1/S期表达上调。I-2/PP 2A过表达抑制细胞增殖.免疫抑制剂的作用机制虽然免疫抑制剂如环孢素A和FK 506的作用靶点被确定为钙调神经磷酸酶，但钙调神经磷酸酶活性的测定方法尚未建立。建立了高效、准确的测定钙调神经磷酸酶活性的方法.翻译与主要信号通路之间的相互作用我们已经鉴定了新的负调控蛋白MKP-7（MAPK途径）和PTP-1（JAK-STAT），并对这些蛋白的作用方式进行了研究。

项目成果

Shinya Mitsuhashi: "Tautomycetin is a novel and specific inhibitor of serine/threonine protein phosphatase type1, PP1"Biochemical and Biophysical Research Communications. 287(2). 328-331 (2001)

Shinya Mitsuhashi：“互变霉素是一种新型、特异性的丝氨酸/苏氨酸蛋白磷酸酶 1 型 PP1 抑制剂”《生物化学和生物物理研究通讯》。

Nobuhiro Tanuma: "Protein tyrosine phosphatase εC selectively inhibits interleukin-6- and interleukin- 10-induced JAK-STAT signaling"Blood. 98(10). 3030-3034 (2001)

Nobuhiro Tanuma：“蛋白质酪氨酸磷酸酶 εC 选择性抑制白细胞介素 6 和白细胞介素 10 诱导的 JAK-STAT 信号”Blood。

Bettina Linck: "Functional properties of the rat phosphatase 1α promoter"Molecular and Cellular Biochemistry. 223(1-2). 123-129 (2001)

Bettina Linck：“大鼠磷酸酶 1α 启动子的功能特性”《分子和细胞生物化学》223(1-2) (2001)。

Nobuhiro Tanuma: "Protein tyrosine phosphatase, PTPεC, inhibits Jak-STAT signaling and differentiation Induced by IL-6 and LIF in M1 leukemia cells"Journal of Biological Biochemistry. 275(36). 28216-28221 (2000)

Nobuhiro Tanuma：“蛋白质酪氨酸磷酸酶，PTPεC，抑制 M1 白血病细胞中 IL-6 和 LIF 诱导的 Jak-STAT 信号传导和分化”《生物化学杂志》275(36)。

Chikako Fukukawa: "Up-regulation of I-2(PP2A)/SET gene expression in rat primary hepatomas and regenerating livers"Cancer Letters. 161(1). 89-95 (2000)

Chikako Fukukawa：“大鼠原发性肝癌和再生肝脏中 I-2(PP2A)/SET 基因表达的上调”癌症快报。