Functional regulation of centrosome and Golgi apparatus by signal-anchoring proteins.

信号锚定蛋白对中心体和高尔基体的功能调节。

• 批准号: 17370049
• 负责人: ONO Yoshitaka
• 金额: $ 10.16万
• 依托单位: Kobe University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-17370049/
• 关键词: Protein kinases; PKN; anchoring proteins; CG-NAP; centrosome; Golgi apparatus

CG-NAP (centrosome and Golgi localized PKN-associated protein) is a coiled-coil protein identified as a binding protein for the regulatory domain of PKN that has a catalytic domain highly homologous to PKC in the carboxyl-terminal region and a unique regulatory domain in the amino-terminal region. In this study, we have analyzed the function of CG-NAP in the Golgi formation and centrosome splitting and obtained the results as follows.1. CG-NAP was found to interact with both microtubules and a cytoplasmic dynein subunit p150^<Glued> and localize to the Golgi apparatus in a microtubule-dependent manner. By examining the recovery process from the depolymerizing microtubules or inhibiting cytoplasmic dynein, it was revealed that CG-NAP is recruited to the minus ends of microtubules by interacting with cytoplasmic dynein, thereby localizes to the Golgi apparatus.2. The Golgi apparatus in mammalian cells forms a continuous ribbon of interconnected stacks of flat cisternae that are positione … More d close to the centrosome. Neither the molecular requirements for, nor the purpose of, Golgi ribbon formation are known. It was revealed that the Golgi apparatus is fragmented in the cells lacking CG-NAP at the Golgi. In these cells transport and glycosylation of membrane proteins occurred normally with some delay, indicating that these stacks are functional. These results suggest that CG-NAP is required for the lateral fusion of the Golgi stacks to form fully functional apparatus.3. The centrosome splitting occurs between duplicated centrosomes at late G2 phase, which is caused by phosphorylation of cohesion proteins between two centrosomes by a protein kinase Nek2A. The ratio of the cells with split centrosomes was increased when CG-NAP or kendrin was suppressed by siRNA. Further, these proteins were found to associate specifically with hyper-phosphorylated inactive Nek2A, suggesting their role in the suppression of Nek2A activity at centrosomes. Possible mechanisms of Nek2A inhibition, such as phosphorylation and binding, are being examined. Less

CG-NAP（中心体和高尔基体定位 PKN 相关蛋白）是一种卷曲螺旋蛋白，被鉴定为 PKN 调节结构域的结合蛋白，其在羧基末端区域具有与 PKC 高度同源的催化结构域，在氨基末端区域具有独特的调节结构域。本研究分析了CG-NAP在高尔基体形成和中心体分裂中的功能，得到如下结果： 1．发现 CG-NAP 与微管和细胞质动力蛋白亚基 p150^<Glued> 相互作用，并以微管依赖性方式定位于高尔基体。通过研究解聚微管或抑制细胞质动力蛋白的恢复过程，发现CG-NAP通过与细胞质动力蛋白相互作用而被募集到微管的负端，从而定位于高尔基体。2．哺乳动物细胞中的高尔基体形成一条连续的带状，由相互连接的扁平池组成，这些池位于靠近中心体的位置。高尔基带形成的分子要求和目的均未知。结果表明，高尔基体缺乏 CG-NAP 的细胞中高尔基体破碎。在这些细胞中，膜蛋白的运输和糖基化正常发生，但有一定的延迟，表明这些堆叠是有功能的。这些结果表明CG-NAP是高尔基体堆叠横向融合形成功能齐全的装置所必需的。3．中心体分裂发生在 G2 晚期的重复中心体之间，这是由蛋白激酶 Nek2A 磷酸化两个中心体之间的粘合蛋白引起的。当CG-NAP或kendrin受到siRNA抑制时，中心体分裂的细胞比例增加。此外，这些蛋白质被发现与过度磷酸化的失活 Nek2A 特异性相关，表明它们在抑制中心体 Nek2A 活性中发挥作用。 Nek2A 抑制的可能机制（例如磷酸化和结合）正在研究中。较少的

项目成果

Formation of morphologically similar globular aggregates from diverse aggregation-prone proteins in mammalian cells.

• DOI: 10.1073/pnas.0409283102
• 发表时间: 2005-08
• 期刊: Proceedings of the National Academy of Sciences of the United States of America
• 影响因子: 11.1
• 作者: H. Mukai;T. Isagawa;Emiko Goyama;Shuhei Tanaka;N. Bence;A. Tamura;Y. Ono;R. Kopito

ラット下垂体由来GH細胞においてTRH刺激で活性化されたPKCεはケラチン8のSer8とSer23をリン酸化する

TRH刺激激活的PKCε磷酸化大鼠垂体来源的GH细胞中角蛋白8的Ser8和Ser23

• 发表时间: 2007
• 作者: Miyamoto;M.;志賀一博;高橋美樹子;秋田朗子
• 通讯作者: 秋田朗子

Silencing of Constitutive NF-kB and JNK Signaling by PKN1 via Phosphorylation of TRAF1

PKN1 通过 TRAF1 磷酸化沉默组成型 NF-kB 和 JNK 信号传导

• 发表时间: 2006
• 作者: Kato;T
• 通讯作者: T

Protein kinase Cs phosphorylates keratin 8 at Ser8 and Ser23 in GH4C1 cells stimulated by thyrotropin-releasing hormone

促甲状腺素释放激素刺激的 GH4C1 细胞中，蛋白激酶 Cs 磷酸化角蛋白 8 Ser8 和 Ser23

• 发表时间: 2007
• 期刊: FEBS J 274
• 作者: Akita;Y
• 通讯作者: Y

Phosphorylation and up-regulation of diacylglycerol kinasey via its interaction with protein kinase C_γ

二酰甘油激酶通过与蛋白激酶 C_γ 的相互作用进行磷酸化和上调

• 发表时间: 2006
• 期刊: J. Biol. Chem 281
• 作者: Yamaguchi;Y
• 通讯作者: Y