New methods for expression cloning of alternative splicing factors

1999 Fiscal Year Final Research Report Summary

• Project/Area Number: 10558102
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 展開研究
• Research Field: Functional biochemistry
• Research Institution: TOKYO MEDICAL & DENTAL UNIVERSITY
• Principal Investigator: HAGIWARA Masatoshi MEDICAL RESEARCH INSTITUTE, TOKYO MEDICAL & DENTAL UNIVERSITY, PROFESSOR, 難治疾患研究所, 教授 (10208423)
• Co-Investigator(Kenkyū-buntansha): MURO Yoshinao NAGOYAMEDICAL UNIVERSITY-ASSISTANT PROFESSOR, 医学部, 助教授 (80270990)
• Project Period (FY): 1998 – 1999
• Keywords: alternative splicing / GFP / arginine methylation / RNAi / SRPK / phosphorylation of SR proteins

Research Abstract

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.

Research Products

• [Publications] Nagai Y, Miyazaki M, Aoki R, Zama T, Inoue S, Hirose K, Iino M, Hagiwara M: "Detection of cAMP-induced phosphorylation with a novel fluorescent indicator."Nat.Biotechnol.. (in press). (2000)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Sakaguchi H, Wada K, Naekawa M Watsuji T, Hagiwara M: "Song Induced Phosphorylation of cAMP Response Element Binding Protein in the song bird brain."J.Neurosciense. 19. 3973-3981 (1999)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Koizumi J, Okamoto Y, Onogi H, Maekawa A, Krainer A, 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)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Okamoto Y, Onogi H, Honda R Hagiwara M et.al: "Cdc2 kinasa-mediated phosphorylation of splicing factor. SF2/ASF."Biochemical and Biophysical Research Communication. 249. 872-878 (1998)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] 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)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Kuroyanagi N, Onogi H, Wakabayashi T, Hagiwara M: "Novel SR-protein-specific kinase, SRPK2, disassembles nuclear speckles."Biochem.Biophys.Res.Commun.. 242. 357-364 (1998)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Nagai, Y., Miyazaki, M., Aoki, R., Zama, T., Inoue, S., Hirose, K., Iino, M., and Hagiwara, M.: "Detection of cAMP-induced phosphorylation with a novel fluorescent indicator."Nat.Biotechnol.. 18. 313-316 (2000)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] 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)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] 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)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] 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)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Muro, Y., Ogawa, Y., Kato, Y., and Hagiwara, M.: "Autoantibody to thioredoxin reductase in an ovarian cancer patient."Biochem.Biophys.Res.Commun.. 242. 267-271 (1998)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Kuroyanagi, N., Onogi, H., Wakabayashi, T., and Hagiwara, M.: "Novel SR-protein-specific kinase, SRPK2, disassembles nuclear speckles."Biochem.Biophys.Res.Commun.. 242. 357-364 (1998)
  • Description: 「研究成果報告書概要(欧文)」より