Study for the cellular signal transduction of Neurofibromatosis Type 1 and Type 2 tumor suppressor gene products to establish the medical molecular target for NF1 and NF2

研究1型和2型神经纤维瘤病抑癌基因产物的细胞信号转导，建立NF1和NF2的医学分子靶点

基本信息

批准号：
16390413
负责人：
ARAKI Norie
金额：
$ 8.77万
依托单位：
Kumamoto University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2004
资助国家：
日本
起止时间：
2004 至 2005
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-16390413/
关键词：
Neurofibromatosis Type 1 NF1 Neurofibromin Neurofibromatosis Type 2 NF2 Merlin Proteomics brain tumor 腫瘍抑制遺伝子

项目摘要

The neurofibromatosis type 1 (NF1) and type 2 (NF2) are autosomal dominantly inherited disorders, and strongly associated with development of intracranial tumors including bilateral vestibular schwannomas, meningiomas and gliomas. To elucidate the biological function of NF1 and NF2 gene products, and, we analyzed their cellular signal transductions by using siRNA techniques and proteomic strategies. 1)NF1 ; NF1 siRNA induces characteristic morphological changes, such as excessive actin stress fiber formation, with elevated phosphorylation levels of coffin, which regulates actin cytoskeletal reorganization by depolymerizing and severing actin filaments, and that was promoted by the activation of a Rho-ROCK-LIMK2 pathway, which requires Ras activation. Fifty three proteins associating to neurofibromin were identified with the quantitative differential proteomic strategy (iTRAQ). These proteins contain collapsin response mediator protein-2 (CRMP-2), calmodulin (CaM), 14-3-3 proteins, Micr … More otubule-associated proteins, coffilin etc. In PC12 cells, the associations of CRMP2 and tubulin to neurofibromin were regulated by their phosphorylation manners. NF1 SiRNA significantly inhibited the neurite extension of PC12 cells, which was correlated with the regulation of CRMP-2 phosphorylations via GSK-3b, Rho-kinase, and other kinases. Neurofibromin has significant roles in the machinery regulating cellular cytoskeletal reorganization, which affects cell motility, adhesion, and differentiations. 2)NF2 ; 28 cellular binding proteins of merlin were identified, and focused on DNA repair enzymes such as poly ADP-ribose polymerase, DNA-PK subunits Ku-antigen 85 and 70. The N-terminal (19-339) region of merlin is essential for their interaction. Subcellular co-localizations of merlin and PARP were observed mainly in their nuclear region with a nuclear export signal (NES) dependent manner of merlin. This nuclear accumulation increased with cellular DNA damages, whereas MEF (PARP-/-) could not accumulate merlin in their nuclear region. These results suggest that merlin is a cytoplasmic-nuclear shuttling protein directed by its NES-dependent transport pathway being associated with PARP. Functional regulations of NF proteins via their cellular associating proteins are essential, and their abnormal regulations caused by NF gene aberrations may be implicated in NF-related pathogenesis. Less

1型神经纤维瘤病（NF1）和2型（NF2）是常染色体，主要遗传性疾病，并且与包括双侧前庭schwannomas，脑膜瘤和胶质瘤在内的颅内肿瘤的发展密切相关。为了阐明NF1和NF2基因产物的生物学功能，并使用siRNA技术和蛋白质组学策略分析了它们的细胞信号转导。 1）NF1 siRNA诱导特征形态的变化，例如过度肌动蛋白应激纤维形成，棺材的磷酸化水平升高，从而调节肌动蛋白细胞骨骼骨骼重组，通过降低和切断肌动蛋白丝的分解，这是通过Rho-Rock-limk2 Pathway的激活来促进的。用定量差异蛋白质组学策略（ITRAQ）鉴定了与神经纤维蛋白相关的53种蛋白质。这些蛋白质含有折叠蛋白反应介质蛋白-2（CRMP-2），钙调蛋白（CAM），14-3-3蛋白，微蛋白，Micro…在PC12细胞中与Otubule相关的蛋白质，咖啡蛋白等，CRMP2和crmp2和小管蛋白与神经纤维蛋白与神经纤维蛋白的关联在PC12细胞中受其磷酸化剂的调节。 NF1 siRNA显着抑制了PC12细胞的神经蛋白延伸，这与通过GSK-3B，Rho-激酶和其他激酶的CRMP-2磷酸化的调节相关。神经纤维蛋白在机械调节细胞骨骼重组中具有重要作用，从而影响细胞运动，粘合剂和分化。 2）NF2;鉴定了28个Merlin的细胞结合蛋白，并集中于DNA修复酶，例如聚ADP-ribose聚合酶，DNA-PK亚基KU-抗原85和70。Merlin的N末端（19-339）区域对于它们的相互作用至关重要。 Merlin的核出口信号（NES）依赖性方式主要在其核区域观察到Merlin和PARP的亚细胞共定位。这种核积累随细胞DNA损伤而增加，而MEF（PARP - / - ）无法在其核区域积聚。这些结果表明，Merlin是一种由NES依赖性转运途径与PARP相关的细胞质核穿梭蛋白。 NF蛋白通过其细胞缔合蛋白的功能调节是必不可少的，其由NF基因畸变引起的异常调节可能与NF相关的发病机理有关。较少的

项目成果

期刊论文数量（31）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

AM-3K, an anti-macrophage antibody, recognizes CD163, a molecule associated with an anti-inflammatory macrophage phenotype

DOI：
10.1369/jhc.5a6871.2006
发表时间：
2006-07-01
期刊：
JOURNAL OF HISTOCHEMISTRY & CYTOCHEMISTRY
影响因子：
3.2
作者：
Komohara, Yoshihiro;Hirahara, Junko;Takeya, Motohiro
通讯作者：
Takeya, Motohiro

Methods for protein purification from biological samples.

从生物样品中纯化蛋白质的方法。

DOI：
发表时间：
2004
期刊：
Tanpakushitsu Kakusan Koso. 49(11)
影响因子：
0
作者：
Kurihara N;Reddy SV;Araki N;Ishizuka S;Ozono K;Cornish J;Cundy T;Singer FR;Roadman GD.;Araki N^*.
通讯作者：
Araki N^*.

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