| Project/Area Number |
10680604
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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 |
Functional biochemistry
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| Research Institution | FUKUI MEDICAL UNIVERSITY |
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Principal Investigator |
INUZUKA Manabu FUKUI MEDICAL UNIVERSITY, SCHOOL OF MEDICINE, ASSOCIATE PROFESSOR, 医学部, 助教授 (00135104)
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| Co-Investigator(Kenkyū-buntansha) |
KIKAWA Yoshiharu FUKUI MEDICAL UNIVERSITY, UNIVERSITY HOSPITAL, LECTURER, 医学部・附属病院, 講師 (90143940)
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| Project Period (FY) |
1998 – 1999
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| Project Status |
Completed (Fiscal Year 1999)
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| Budget Amount *help |
¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 1999: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1998: ¥2,000,000 (Direct Cost: ¥2,000,000)
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| Keywords | GLUCONEOGENESIS / FRUCTOSE-1,6-BISPHOSPHATASE / TRANSCRIPTIONAL REGULATION / VITAMIN D3 / RETINOIC ACID / NUCLEAR RECEPTOR PROTEIN / VDRE / RARE / GENE DIAGNOSIS / VDRE |
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| Research Abstract |
Fructose-1,6-bisphosphatase (FBPase) is a key gluconeogenic enzyme. We found that both enzyme activity and mRNA level of the human FBPase gene are increased by 9-cis-retinoic acid (9cRA) and all-trans-retinoic acid (atRA) as well as 1,25-dihydroxyvitamin D3 (VD3) in human promyelocytic HL60 cells and normal monocytes in peripheral blood which we have used as an alternative source to liver samples for DNA diagnosis of FBPase deficiency. To understand the mechanism of this molecular action, the 2.4 kb 5'-regulatory region of human FBPase gene was cloned and sequenced. Using cotransfection assays in CV-1 cells, a 0.5 kb FBPase basal promoter fragment was found to confer induction by VD3, 9cRA and atRA, which was respectively mediated by vitamin D3 receptor (VDR), retinoid X receptor (RXR), and retinoic acid receptor (RAR). Within this region, we identified a direct repeat sequence, 5'-TAACCTttcTGAACT-3' (-340 to -326), which functioned as a common response element for VD3, 9cRA and atRA. Results of electrophoretic mobility shift assays indicated that VDR-RXR and RAR-RXR heterodimers specifically bind this response element. These observations suggest that VD3 and RA are important modulators of the human FBPase gene expression in the monocytic blood cells.
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