Study on cellular signal transduction of Neurofibromatosis Type 1 and Type 2 tumor suppressor gene products

神经纤维瘤病1型和2型抑癌基因产物的细胞信号转导研究

基本信息

批准号：
14571319
负责人：
ARAKI Norie
金额：
$ 2.56万
依托单位：
Kumamoto University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2003
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14571319/
关键词：
Neurofibromatosis Type 1 NF1 Neurofibromin Neurofibromatosis Type 2 NF2 Merlin Proteomics brain tumor プロテミクス脳腫瘍学習障害

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. NF1 and NF2 genes were recently identified on the different locus, and the proteins they encode (termed as neurofibromin for NF1 and merlin for NF2) were found to be no homologies despite of their pathological similarity. To elucidate the biological function of NF1 and NF2 proteins, we analyzed their signal transductions by cellular proteomic and biological strategies. 1)NF1 : Neurofibromin has a domain acting as a GAP and thought to be an important Ras regulator. We identified 35 novel cellular neurofibromin-associating proteins. 14-3-3 was identified as one of the most interesting core candidates of NF1 regulators. The interaction of 14-3-3 is mainly directed to the C-terminal domain (CTD) of neurofibromin, and the cAMP-dependent protein kinase (PKA)-depend … More ent phosphorylation clustered on CTD-Ser (2576, 2578, 2580, 2813) and Thr (2556) is required for the interaction. Interestingly, the increased phosphorylation and association of 14-3-3 negatively regulate the GAP activity of neurofibromin. These findings indicate that PKA phosphorylation followed by 14-3-3 protein and other protein cluster interactions may modulate the biochemical and biological functions of cellular neurofibromin. 2)NF2 ; Cellular binding proteins of merlin were identified as poly ADP-ribose polymerase, DNA-PK subunits Ku-antigen 85 and 70 by MAS spec analysis. 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 PARP gene deficient MEF could not accumulate merlin in their nuclear region, even after the treatments of LMB or bleomycin, These results suggest that merlin is a cytoplasmic-nuclear shuttling protein directed by its NES-dependent transport pathway being associated with PARP. The tumor suppressive function of merlin may be linked to cellular DNA-repair and related signals via the interaction of cellular merlin binding proteins such as PARP and DNA-PKs. 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。最近在不同的基因座上鉴定了NF1和NF2基因，尽管它们具有病理相似性，但发现它们编码的蛋白质（称为NF1的神经纤维蛋白和NF2的神经纤维蛋白）不是同源性。为了阐明NF1和NF2蛋白的生物学功能，我们通过细胞蛋白质组学和生物学策略分析了它们的信号翻译。 1）NF1：神经纤维蛋白的结构域充当间隙，被认为是重要的RAS调节剂。我们确定了35种新型细胞神经纤维蛋白促蛋白。 14-3-3被确定为NF1调节器中最有趣的核心候选者之一。 14-3-3的相互作用主要针对神经纤维蛋白的C末端结构域（CTD），以及与CAMP依赖的蛋白激酶（PKA）依赖性……更多的ENT磷酸化聚类在CTD-SER上（2576，2576，2578，2578，2580，2813）和THR（25556）和THR（需要）。有趣的是，14-3-3的磷酸化和缔合的增加对神经纤维素的间隙活性负调节。这些发现表明，PKA磷酸化，然后是14-3-3蛋白和其他蛋白质簇相互作用可能会调节细胞神经纤维蛋白的生化和生物学功能。 2）NF2;通过MAS Spec分析，Merlin的细胞结合蛋白被鉴定为Poly ADP-核糖聚合酶，DNA-PK亚基KU抗原85和70。梅林的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。这种核积累会随细胞DNA损伤而增加，而PARP基因缺乏MEF也无法在其核区域中积累MERLIN，即使在LMB或BLEOMYCIN的处理后，这些结果表明Merlin是一种依赖NES依赖性的运输路径与PARP相关的细胞质核核核穿梭蛋白。 Merlin的肿瘤抑制功能可以通过细胞Merlin结合蛋白（如PARP和DNA-PKS）的相互作用与细胞DNA修复和相关信号联系起来。较少的