| Project/Area Number |
17590694
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Gastroenterology
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| Research Institution | Institute for Developmental Research, Aichi Hunan Service Center |
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Principal Investigator |
INAGUMA Yutaka Institute for Developmental Research, Department of Molecular Neurobiology, 神経制御学部, 主任研究員 (10250250)
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| Co-Investigator(Kenkyū-buntansha) |
WATANABE Kimi Institute for Developmental Research, Research Management and Support Service, 研究企画調整科, 研究助手 (50393145)
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| Project Period (FY) |
2005 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2006: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 2005: ¥2,300,000 (Direct Cost: ¥2,300,000)
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| Keywords | SUMO / pancreas / acinar cell / ubiquitin-like protein / TTRAP / UBC9 / FOXP2 / SMAD4 |
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| Research Abstract |
We found that both SUMO-1 and SUMO-2 were present at high levels in pancreatic acinar cells and investigated its significance. TTRAP was isolated as a partner to interact with SUMO-2 when a yeast two-hybrid screening was performed. TTRAP has features as a phosphodiesterase and showed a weak AP-endonuclease-like activity. We further investigated that FOXP2 interacted with TTRAP and UBC9, a specific conjugating enzyme of SUMO. FOXP2 is a transcription factor and is expressed in the various tissues. We confirmed that FOXP2 was present in the pancreas by the Western blot analysis. In the present study, to determine the target site for sumoylation of FOXP2, eukaryotic sumoylation in Escherichia coli was performed. Three sites (K74, K274, and K673) were predicted as candidates to be sumoylated in silico. As a result, when lysine at 673 was substituted by arginine, the sumoylation of FOXP2 was lost. This indicated that K673 was the target residue to be sumoylated. Furthermore, we investigated transcriptional activities of wild type and K673R by using a reporter assay. The reporter gene was driven under the thymidine kinase promotor, and the FOXP2-binding sequence from CC10 gene was inserted into the reporter vector. The repression by K673R was stronger than the one by wild type suggesting that the sumoylation of FOXP2 affected the DNA-binding activity.
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