| Project/Area Number |
11470307
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| Research Category |
Grant-in-Aid for Scientific Research (B).
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Orthopaedic surgery
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| Research Institution | SHINSHU UNIVERSITY, SCHOOL OF MEDICINE |
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Principal Investigator |
EBARA Sohei DEPARTMENT OF ORTHOPAEDIC SUEGERY, SHINSHU UNIVERSITY, SCHOOL OF MEDICINE, ASSSITANT PROFESSOR, 医学部・附属病院, 講師 (40176780)
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| Co-Investigator(Kenkyū-buntansha) |
TAKAOAK Kunio School of Medicine, Shinshu Univ. Professor, 医学部, 教授 (30112048)
NAKAYAMA Kohzo School of Medicine, Shinshu Univ. Lecturer, 医学部, 講師 (70192680)
NIKAIDO Toshio School of Medicine, Shinshu Univ. Lecturer, 大学院・医学研究科, 助教授 (50180568)
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| Project Period (FY) |
1999 – 2000
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| Project Status |
Completed (Fiscal Year 2000)
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| Budget Amount *help |
¥5,400,000 (Direct Cost: ¥5,400,000)
Fiscal Year 2000: ¥2,600,000 (Direct Cost: ¥2,600,000)
Fiscal Year 1999: ¥2,800,000 (Direct Cost: ¥2,800,000)
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| Keywords | BONE MORPHOGENETIC PROTEIN / PROMOTER ACTIVITY / E-BOX / HELIX-LOOP-HELIX / USF / OSTEOBLASTIC CELLS / TRANSCRIPTIONAL REGULATION / 5'-FLANKING REGION / 骨形成 / BMP-4 |
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| Research Abstract |
We analyzed the promoter activity of murine bone morphogenetic protein-4 (mBMP-4) and determined that the 5'-flanking region of exon I is involved mainly in transcriptional regulation and position-265 to-246 of 5'-flanking region of mBMP-4 gene acts as a cis-element important for the regulation of BMP4 transcription in MC3T3E1 cells. By use of site-directed mutagenesis, we have established that the E-box, CACGTG, located within this short region of promoter, is essential for transcriptional activation of the mBMP-4 gene. Upstream stimulatory factor (USF), a member of helix-loop-helix (HLH) group of regulatory proteins, was found to bind to this E-box using supershift gel mobility analysis. We also analyzed the promoter activity of mouse BMP-4 gene in three different types of cell lines (MC3T3E1, ST2 and C6 glioma cells) and demonstrated that position-786 to-691 of the 5'-flanking region of exon I plays a critical role for regulation of tissue specific expression of mouse BMP-4 gene in cells of an osteogenic lineage. By use of site-directed mutagenesis, we have established that the E-box, CATCTG, located within this 5'-flanking region, is essential for tissue specific transcriptional activation of mouse BMP-4 gene and demonstrated that an osteogenic lineage-specific novel transcriptional factor (s) recognizes this E-box. It is proposed that the HLH transcription regulatory proteins play an important role in mBMP-4 gene transcription in this osteoblastic cell line, like that of Myo-D in myogenic differentiation.
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