Molecular cloning and functional analyses of a novel maternal neurogenic gene, neurotic that is essential for Notch signaling.

一种新型母体神经源性基因的分子克隆和功能分析，该基因对 Notch 信号传导至关重要。

• 批准号: 14580717
• 负责人: MATSUNO Kenji
• 金额: $ 2.3万
• 依托单位: Tokyo University of Science
• 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-14580717/
• 关键词: Notch; neurotic; neurogenic gene; 0-fucosyltransferase; 0-fucT1; 0-fucose; Drosophila; development; O-フコース; O-フコース転移酵素; 母性効果; 細胞間相互作用

Notch signalling, which is highly conserved from nematodes to mammals, plays critical roles in many developmental processes.In the Drosophila embryo, deficiency in Notch signalling results in neurohyperplasia, commonly referred to as the neurogenic phenotype.Here we identify a novel maternal neurogenic gene, neurotic, and show that it is essential for Notch signalling, neurotic encodes a protein highly homologous to mammalian GDP-fucose protein O-fucosyltransferase, which adds fucose sugar to epidermal growth factor-like repeats.neurotic functions in a cell autonomous manner, and genetic epistasis tests reveal that Neurotic is required for the activity of full-length but not an activated form of Notch.Further, we show that neurotic is required for Fringe activity, which encodes a fucose-specific,β1,3 N-acetylglucosaminyltransferase, previously shown to modulate Notch receptor activity.Finally, Neurotic is essential for the physical interaction of Notch with its ligand Delta, and for Fringe ability to modulate this interaction in Drosophila cultured cells.We present here an unprecedented example of an absolute requirement of a protein glycosylation event for a ligand-receptor interaction.Our results suggest,that O-fucosylation catalyzed by Neurotic is also involved in the Fringe-independent activities of Notch and may provide a novel on-off mechanism that regulates ligand-receptor interactions.

Notch信号在线虫和哺乳动物中高度保守，在许多发育过程中起着关键作用。在果蝇胚胎中，Notch信号的缺陷导致神经增生，通常被称为神经原性表型。在这里，我们鉴定了一个新的母体神经原性基因neurotic，并表明它对Notch信号至关重要，神经症编码一种与哺乳动物GDP-岩藻糖蛋白O-岩藻糖基转移酶高度同源的蛋白质，其将岩藻糖添加到表皮生长因子样重复序列中。神经症以细胞自主方式发挥功能，和遗传上位性测试表明，神经质是全长Notch活性所必需的，而不是活化形式的Notch。此外，我们表明，神经质是Fringe活性所必需的，Fringe活性编码岩藻糖特异性β 1，3 N-乙酰葡糖胺转移酶，先前显示调节Notch受体活性。最后，神经质对于Notch与其配体Delta的物理相互作用以及Fringe在果蝇培养细胞中调节这种相互作用的能力是必不可少的。我们在这里提出了一个前所未有的例子，说明配体-受体相互作用绝对需要蛋白质糖基化事件。我们的结果表明，Neurotic催化的O-岩藻糖基化也参与了Notch的Fringe非依赖性活性，并可能提供一种新的调节配体-受体相互作用

项目成果

Takahashi, S., Yoshimori, H., Yamasaki, N., Matsuno, K., Murakami, R.: "Cell-fate choice and boundary formation by combined action of Notch and engrailed in the Drosophila hindgut."Dev.Genes Evol.. 212. 534-541 (2002)

Takahashi, S.、Yoshimori, H.、Yamasaki, N.、Matsuno, K.、Murakami, R.：“通过Notch和果蝇后肠中的Notch的联合作用来选择细胞命运和边界形成。”Dev.Genes Evol

Watanabe, N., Tezuka, Y., Matsuno, K., Miyatani, S., Morimura, N., Yasuda, M., Fujimaki, R., Kuroda, K., Hiraki, Y, Hozumi, N., Tezuka, K.: "Suppression of differentiation and proliferation of early chondrogenic cells by Notch."J.Bone Miner.Metab.. 21. 34

渡边，N.，手冢，Y.，松野，K.，宫谷，S.，森村，N.，安田，M.，藤卷，R.，黑田，K.，平木，Y，保住，N.，手冢

Momoko Hase: "Expression and characterization of Drosophila X11-like/Mint protein during neural development"J. Neurochem.. 81. 1223-1232 (2002)

Momoko Hase：“果蝇 X11 样/薄荷蛋白在神经发育过程中的表达和表征”J。

Kenji Matsuno: "Involvement of a proline-rich motif and RING-H2 finger of Deltex in the regulation of Notch signaling"Development. 129. 1049-1059 (2002)

Kenji Matsuno：“Deltex 富含脯氨酸的基序和 RING-H2 指参与 Notch 信号传导的调节”开发。

Sasamura, T: "neurotic, a novel maternal neurogenic gene, encodes an O-fucosyltratisferase that is essential for Notch-Delta interactions."Development. 130. 4785-4795 (2003)

Sasamura，T：“neurotic，一种新型母体神经源性基因，编码一种 O-岩藻糖基转移酶，该酶对于 Notch-Delta 相互作用至关重要。”开发。