2003 Fiscal Year Final Research Report Summary
The analysis of a newly identified molecule which interacts with phosphodiesterase3B insulin-dependently
| Project/Area Number |
14571095
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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 |
Metabolomics
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| Research Institution | Ehime University |
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Principal Investigator |
ONUMA Hiroshi Ehime University, School of Medicine, Instructor, 医学部, 助手 (00294794)
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| Co-Investigator(Kenkyū-buntansha) |
OSAWA Haruhiko Ehime University, School of Medicine, Associate Professor, 医学部, 助教授 (90294800)
MAKINO Hideichi Ehime University, School of Medicine, Professor, 医学部, 教授 (50009578)
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| Project Period (FY) |
2002 – 2003
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| Keywords | Insulin / Phosbodiesterase 3B / Interaction / 14-3-3β / Serine phosphorylation / Adipocytes |
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| Research Abstract |
We have identified 14-3-3β, a critical scaffolding molecule in signal transduction, as a protein that interacts with PDE3B using the yeast two-hybrid system. The interaction between PDE3B and 14-3-3β was then confirmed in vitro. GST pull-down using truncated form of mouse PDE3B reveals that the region of mouse PDE3B that interacts with 14-3-3β is located between a 223 and 322. The GST-tagged 14-3-3β interacts with endogenous PDE3B of rat adipocytes, and this interaction is enhanced when adipocytes are treated with insulin. Coimmunoprecipitation experiments reveal that endogenous PDE3B also associates with endogenous 14-3-3β in rat adipocytes and this interaction is enhanced by insulin. Treatment of adipocytes with 1μM okadaic acid, a potent, cell-permeable Ser/Thr phosphatase inhibitor, increased the association of PDE3B with 14-3-3β to the same level as that achieved with, insulin. Two different PI 3-K-inhibitors, wortmannin and Ly294002 block this induction, suggesting that PI 3-K is required. Synthetic 15 amino acid peptides of rat PDE3B containing phosphorylated Ser-279 or 302 inhibit this interaction, indicating that the insulin-regulated phosphorylation of these serine residues is involved. 14-3-3β could function as a scaffolding protein in the activation of PDE3B by insulin.
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