CHARACTERIZATION OF SIGNAL TRANSDUCTION SYSTEM OF IL-13 RECEPTOR -MOLECULAR CLONING OF A NOVEL SIGNAL TRANSDUCTION MOLECULES FOR IL-13 SIGNALING.

IL-13 受体信号转导系统的表征 - 用于 IL-13 信号转导的新型信号转导分子的分子克隆。

• 批准号: 12672112
• 负责人: MURATA Takashi
• 金额: $ 2.05万
• 依托单位: Nagasaki University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12672112/
• 关键词: IL-13; STATG; TRAF3; IL13RBP1; MIP-T3; signaltransduction; インターロイキン13; アレルギー; Th2; サイトカイン; シグナル伝達; インターロイキン; レセプター; Yeast tri-hybrid

Current known IL-13 signaling is mainly mediated through IL-4Ra, but few are known about IL-13Ra1's downstream. Yeast tri-hybrid system was utilized for searching proteins which can associate with IL-13 receptor. We found a novel IL-13Ra1 binding protein from human fetal cDNA library and named as IL13RBP1 (Genbank Accession Number : A242456). Through northern blot analysis, 2 kinds of IL13RBP1 mRNA, 4.4 kb and 2.4 kb, were detected in all tissues examined. In testis, additional bands were detected between 2.5-2.7 kb. IL13RBP1 gene cloned from human testis cDNA library has a whole length of 2568 by and an open reading frame of 692 aa. While in normal tissues, IL13RBP1 has an open reading frame of 625 aa, which lacks an insert fragment in the middle part compared that in testis. IL13RBP1's association with IL-13Ral was proved in yeast two-, tri-hybrid system and mammalian cells : Interestingly, the association was independent of tyrosine phosphorylation. Besides interaction with IL-13Ra1, IL13RBP1 was found to inhibit STATE tyrosine phosphorylation in response to IL-4 and IL-13 stimulation. Furthermore, STAT6's DNA binding activity and transcriptional activity were also partly inhibited by transient expressed IL13RBP1 Interestingly, IL13RBP1 was found to encode the same protein as MIP-T3 (Microtubule interacted protein that associated with TRAF3). MIP-T3 constitutively interacts with TRAF3 protein. MIP-T3 was found to bind to microtubule and tubulin in vitro. These findings indicate that ILl3RBPl/MIP-T3 may play a role in both IL-13 and CD40 signaling, but detailed mechanism has to be further investigated.Our results suggest that IL13RBP1 is a novel inhibitor of IL-13 signaling and may be a useful molecular in ameliorating various conditions including allergies, pulmonary asthma, parasitic infection and cancer in which IL-13 plays a central role.

目前已知的IL-13信号主要通过IL-4Ra介导，但对IL-13Ra1的下游知之甚少。利用酵母三杂交系统寻找与IL-13受体相关的蛋白。我们从人胎儿cDNA文库中发现了一个新的IL-13Ra1结合蛋白，命名为IL13RBP1 （Genbank登录号：A242456）。通过northern blot分析，在所有检测组织中均检测到4.4 kb和2.4 kb的IL13RBP1 mRNA。在睾丸中，在2.5-2.7 kb之间检测到额外的条带。从人睾丸cDNA文库中克隆的IL13RBP1基因全长2568，开放阅读框692 aa。而在正常组织中，IL13RBP1具有625aa的开放阅读框，与睾丸相比，在中间部分缺少插入片段。在酵母二杂交、三杂交系统和哺乳动物细胞中证实了IL13RBP1与IL-13Ral的关联：有趣的是，这种关联不依赖于酪氨酸磷酸化。除了与IL-13Ra1相互作用外，IL13RBP1在IL-4和IL-13刺激下抑制STATE酪氨酸磷酸化。此外，STAT6的DNA结合活性和转录活性也被瞬时表达的IL13RBP1部分抑制。有趣的是，IL13RBP1被发现编码与MIP-T3（与TRAF3相关的微管相互作用蛋白）相同的蛋白质。MIP-T3与TRAF3蛋白组成性相互作用。在体外发现MIP-T3与微管和微管蛋白结合。这些发现表明，ILl3RBPl/MIP-T3可能在IL-13和CD40信号传导中都起作用，但详细的机制有待进一步研究。我们的研究结果表明，IL13RBP1是一种新的IL-13信号抑制剂，可能是一种有用的分子，可以改善各种疾病，包括过敏、哮喘、寄生虫感染和癌症，其中IL-13起着核心作用。

项目成果

Niu Y, Murata T, Waianabe K, Kawakami K, Yoshimura A, Inoue J, Puri RK, Kobayashi N: "MIP-T3 Associates with IL-13Rα1 and Suppresses STAT6 Activation in Response to IL-13 Stimulation."FEES letters. (In press). (2003)

Niu Y、Murata T、Waianabe K、Kawakami K、Yoshimura A、Inoue J、Puri RK、Kobayashi N：“MIP-T3 与 IL-13Rα1 相关并抑制 STAT6 激活以响应 IL-13 刺激。”费用信件（。 (2003)

Niu Y, Murata T, Watanabe K, Kawakami K, Yoshimura A, Inoue J, Puri RK, Kobayashi N: "MIP-T3 Associates with IL-13Rα1 and Suppresses STAT6 Activation in Response to IL-13 Stimulation"FEBS letters. (In press). (2003)

Niu Y、Murata T、Watanabe K、Kawakami K、Yoshimura A、Inoue J、Puri RK、Kobayashi N：“MIP-T3 与 IL-13Rα1 相关并抑制 STAT6 激活以响应 IL-13 刺激”FEBS 信件（2003 年）。 ）

Niu Y, Murata T, Watanabe K, Kawakami K, 'Yoshimura A, Inoue J, Puni RK, Kobayashi N.: "MIP-T3 Associates with IL-13Rα1 and Suppresses STATG Activation in Response to IL-13 Stimulation"FGBSLCTTCRS. (in press).

Niu Y、Murata T、Watanabe K、Kawakami K、Yoshimura A、Inoue J、Puni RK、Kobayashi N.：“MIP-T3 与 IL-13Rα1 相关并抑制 STATG 激活以响应 IL-13 刺激”FGBSLCTTCRS。按）。