| Project/Area Number |
04671411
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
応用薬理学・医療系薬学
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| Research Institution | Osaka University |
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Principal Investigator |
OSUGI Takeshi Osaka University School of Medicine Assistant Prof.Faculty of Pharmaceutical, 医学部, 助手 (50176880)
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| Project Period (FY) |
1992 – 1993
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| Project Status |
Completed (Fiscal Year 1993)
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| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1993: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1992: ¥1,300,000 (Direct Cost: ¥1,300,000)
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| Keywords | Drug Tolerance / Drag Dependence / Morphine / Single-stranded DNA / DNA Binding Protein / cAMP Response Element / Mouse / Cerebellum / 薬物耐性依存 / 遺伝子発現 / cAMP response ehement(CRE) |
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| Research Abstract |
It has been speculated that opiate tolerance and dependence may occur at the level of gene expression. We found that the binding activity of a nuclear factors (ssCRE-BP) to single-stranded CRE of somatostatin gene is altered by long-term treatment with morphine in the mouse cerebellum. To understand the function of ssCRE-BP, the protein was purified from the mouse cerebellum by a combinationof chromatography on DNA affinity agarose and MONO Q HR.The native protein exhibited a molecular size of 110-150 kDa by gel filtration and two polypeptides of about 35-40 kDa were observed on SDS-PAGE.The purified ssCRE-BP had low binding activity, but the activity was recovered in the presence of flow-through fractions of DNA affinity chromatography. The binding activity of purified ssCRE-BP was also increased by the additionof casein but not by BSA and histone. To investigate that ssCRE-BP is different from CREBs, antisera raised against 43 kDa-CREB or CRE-BPI were used in gel shift assay. The antisera did not interact with ssCRE-BP in the assay. Lysyl endopeptidase-digested peptides of ssCRE-BP were prepared and purified by reverse-phase HPLC.Some of the peptides were suquenced by automated Edman degration. Database searches showed low homology to other known protein from mammalian orgin. These data indicate that ssCRE-BP is a novel CRE-binding protein whose binding activity is increased by casein-like factors.
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