New methods for expression cloning of alternative splicing factors

选择性剪接因子表达克隆的新方法

• 批准号: 10558102
• 负责人: HAGIWARA Masatoshi
• 金额: $ 8.7万
• 依托单位: TOKYO MEDICAL & DENTAL UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10558102/
• 关键词: alternative splicing; GFP; arginine methylation; RNAi; SRPK; phosphorylation of SR proteins; カルシトニン; alternative splicing; RT-PCR; SF2; ASF; リン酸化反応; スプライシング

We tried to establish a new method to screen RNA splicing factors using a calcitonin/CGRP mini gene with GFP expression vector. Although we had expected that regulators for neuron-specific alternative splicing could be cloned with this system, no change of GFP expression was observed in any cells used for this experiment. However, we succeeded to get clones of potential RNA-binding proteins with another method which we originally developed.

我们试图建立一种新方法，使用具有GFP表达载体的降钙素/CGRP MINI基因筛选RNA剪接因子。尽管我们期望通过该系统克隆用于神经元特异性替代剪接的调节剂，但在用于该实验的任何细胞中均未观察到GFP表达的变化。但是，我们成功地通过我们最初开发的另一种方法获得了潜在的RNA结合蛋白的克隆。

项目成果

Sakaguchi, H., Wada, K., Maekawa, M., Watsuji, T., and Hagiwara, M.: "Song-Induced Phosphorylation of cAMP Response Element Binding Protein in the Songbird Brain."J.Neuroscience.. 19. 3973-81 (1999)

Sakaguchi, H.、Wada, K.、Maekawa, M.、Watsuji, T. 和 Hagiwara, M.：“鸣禽大脑中 cAMP 响应元件结合蛋白的歌曲诱导磷酸化。”J.Neuroscience.. 19。

Okamoto, Y., Onogi, H., Honda, R., Yasuda, H., Wakabayashi, T., Nimura, Y., and Hagiwara, M.: "cdc2 kinasa-mediated phosporylation of splicing factor. SF2/ASF."Biochemical and Biophysical Research Communications.. 249. 872-878 (1998)

Okamoto, Y.、Onogi, H.、Honda, R.、Yasuda, H.、Wakabayashi, T.、Nimura, Y. 和 Hagiwara, M.：“cdc2 kinasa 介导的剪接因子磷酸化。SF2/ASF。

Muro Y, Ogawa Y, Kato Y Hagiwara M: "Autoantibody to thioredoxin reductase in an ovan an cancer patient."Biochem.Biophys.Res.Commun.. 242. 267-271 (1998)

Muro Y、Okawa Y、Kato Y Hagiwara M：“癌症患者卵巢中硫氧还蛋白还原酶的自身抗体。”Biochem.Biophys.Res.Commun.. 242. 267-271 (1998)

Kuroyanagi Noriyo,Onogi Hiroshi,Wakabayashi Takashi,Hagiwara M.: "Novel SR-protein-specific kinase,SRPK2,disassembles nuclear speckles." Biochem.Biophys.Res.Commun.242,. 357-364 (1998)

Kuroyanagi Noriyo、Onogi Hiroshi、Wakabayashi Takashi、Hagiwara M.：“新型 SR 蛋白特异性激酶 SRPK2，可分解核斑点。”

Koizumi, J., Okamoto, Y., Onogi, H., Mayeda, A., and Krainer, A., and Hagiwara, M.: "The subcellular localization of SF2/ASF is regulated by the direct interaction with SR protein kinases (SRPKs)."J.Biol.Chem.. 274. 11125-11131 (1999)

Koizumi, J.、Okamoto, Y.、Onogi, H.、Mayeda, A.、Krainer, A. 和 Hagiwara, M.：“SF2/ASF 的亚细胞定位是通过与 SR 蛋白激酶的直接相互作用来调节的