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 型神经纤维瘤病 (NF2) 是常染色体显性遗传性疾病，与颅内肿瘤（包括双侧前庭神经鞘瘤、脑膜瘤和神经胶质瘤）的发展密切相关，最近在不同基因座上发现了 NF1 和 NF2 基因。他们编码的蛋白质（对于 NF1 称为神经纤维蛋白，对于 NF2 称为 merlin）被发现没有同源性，尽管它们具有病理性为了阐明 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转导。 1)NF1：神经纤维蛋白具有充当 GAP 的结构域，并且被认为是重要的 Ras 调节因子。 14-3-3 被认为是最有趣的 NF1 调节因子核心候选蛋白之一。 14-3-3 的相互作用主要针对 NF1 调节因子。神经纤维蛋白的 C 末端结构域 (CTD) 和 cAMP 依赖性蛋白激酶 (PKA) 依赖性…更多 ent 磷酸化聚集在 CTD-Ser (2576, 2578, 2580, 2813) 和 Thr (2556) 上，是相互作用所必需的暗示地，14-3-3 磷酸化和关联的增加负向调节神经纤维蛋白的 GAP 活性。这些发现表明 PKA。 14-3-3 蛋白和其他蛋白簇相互作用的磷酸化可能调节细胞神经纤维蛋白的生化和生物学功能；merlin 的细胞结合蛋白被鉴定为聚 ADP-核糖聚合酶、DNA-PK 亚基 Ku 抗原。 85 和 70 通过 MAS 规范分析，merlin 的 N 末端 (19-339) 区域对于它们的亚细胞共定位至关重要。 merlin 和 PARP 主要在其核区域中观察到，merlin 的核输出信号 (NES) 依赖性方式随着细胞 DNA 损伤而增加，而 PARP 基因缺陷的 MEF 即使在处理后也无法在其核区域中积累 merlin。这些结果表明，merlin 是一种细胞质-核穿梭蛋白，由其与 PARP 相关的 NES 依赖性转运途径引导。merlin 的肿瘤抑制功能可能与细胞有关。通过细胞 merlin 结合蛋白（如 PARP 和 DNA-PK）的相互作用进行 DNA 修复和相关信号。