| Project/Area Number |
05557009
|
|---|
| Research Category |
Grant-in-Aid for Developmental Scientific Research (B)
|
| Allocation Type | Single-year Grants |
|---|
| Research Field |
General pharmacology
|
|---|
| Research Institution | Osaka University |
|---|
Principal Investigator |
MIKI Naomasa Osaka Univ.Faculty of Med., Professor, 医学部, 教授 (40094445)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
YANAGITA Tomoji Central Insti.Exp.Animals, Director, 研究所長
KUO Che-hui Osaka Univ.Faculty of Med., Assis.Prof., 医学部, 助手 (50126570)
OSUGI Takeshi Osaka Univ.Faculty of Med., Assis.Prof., 医学部, 講師 (50176880)
HIGUCHI Hiroshi Osaka Univ.Faculty of Med., Assoc.Prof., 医学部, 助教授 (30150337)
渡辺 康裕 防衛医科大学校, 教授 (90127324)
王 小兵 大阪大学, 医学部, 助手 (30243207)
|
|---|
| Project Period (FY) |
1993 – 1995
|
| Project Status |
Completed (Fiscal Year 1995)
|
| Budget Amount *help |
¥13,000,000 (Direct Cost: ¥13,000,000)
Fiscal Year 1995: ¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1994: ¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 1993: ¥7,400,000 (Direct Cost: ¥7,400,000)
|
|---|
| Keywords | morphine / tolerance / dependence / ssCRE-BP / puraalpha / DNA-binding protein / cAMP response element / casein / CAMPレスポンスエレメント / 一本鎖CRE結合蛋白 / 転写因子 / cDNAクローニング / 大脳 / 小脳 / DNA結合蛋白 / ISDB法 / メタンフェタミン / 薬物依存 |
|---|
| Research Abstract |
Chronic administration of morphine to NG108-15 cells or mice decreased the binding activity of a nuclear factor (ssCRE-BP) tosingle-stranded oligo-DNA of cAMP-response element (ssCRE). we have purified ssCRE-BP from mouse cerebella and isolated the cDNA clone. We found that ssCRE-BP from mouse brain was identical to Pura, which is a binding protein for single-stranded purine-rich DNA sequences that was originally isolated as a replication factor. ssCRE-BP was abundant in the cerebral cortex and cerebellum, but very low levels were detected in other tissues. Examination of the effect of chronic morphine treatment on the amount of ssCRE-BP in mouse brain showed that ssCRE-BP was not influenced by chronic morphine treatment significantly in western blot analysis. Although the DNA binding activities of purified ssCRE-BP or its GST-fusion protein were very low, they were markedly enhanced by the addition of an activator that was about 100kDa, trypsin sensitive and heat stable. The activator can be substituted with casein, which increased the affinity of ssCRE-BP for DNA by one order. A series of deletion mutants were prepared for determining the DNA binding-and activator-interacting domains and both of them were found to reside in AA 50-215 of ssCRE-BP.Although ssCRE-BP was abundant in the brain, the activator was ubiquitous. Characterization of the activator may provide insight into the physiological roles of ssCRE-BP which is involved in the development of morphine dependence and tolerance.
|
