| Project/Area Number |
04680195
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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 |
代謝生物化学
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| Research Institution | The University of Tokushima |
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Principal Investigator |
YOSHIOKA Hidefumi The University of Tokushima, Faculty of Engineering, Assistant Professor, 工学部, 助教授 (40191548)
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| Co-Investigator(Kenkyū-buntansha) |
OHUCHI Hideyo The University of Tokushima, Faculty of Engineering, Assistant, 工学部, 助手 (00253229)
NOJI Sumihare The University of Tokushima, Faculty of Engineering, Professor, 工学部, 教授 (40156211)
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| Project Period (FY) |
1992 – 1994
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| Project Status |
Completed (Fiscal Year 1994)
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| Budget Amount *help |
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1994: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 1993: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 1992: ¥1,400,000 (Direct Cost: ¥1,400,000)
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| Keywords | P-450 / Growth Hormone / Male-specific expression / steroid 16alpha hydroxylase / Wnt / チトクロームP-450 / シス-エレメント / 置伝子発現 |
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| Research Abstract |
Sex-specific P-450s are all steroid hydroxylases and have evolved as members within the differnt subfamilies. Mouse P-45016alpha is the male -specific steroid 16alpha-hydroxylase, while P450cb (the other IID subfamily-member) does not catalyze metabolism of steroids and is not sex-specific in its expression. I performed DNase I footprinting and in vitro transcription with the 5'-flanking sequence of P-45016alpha gene and compared them with those seen in the structurally related P450cb gene. As a result, two cis-acting transcription elements were identified : SDI (sex different information) and CTE (comon transcription element). The former is a strong element specific for the P-45016alpha gene and is located in the -84/-102, while the latter element is weak and is present in the -44/-67 of both P-45016a and P450cb genes. Futhermore, I purified a SDI binding protein from liver nuclear extract of male mouse using a heparin-agarose colume, DEAE cellurose colume, and specific -DNA affinity colume. This SDI-binding protein was determined its partial amino acid sequence and deganarate oligonucleotide was used for a probe of cDNA cloning experiment. I isolated several positive clones. A complete structure of cDNA will be evaluated. The p-45016a gene promoter, however, does not elicit the male-specific in vitro transcription, indicating that the cis-acting elements other than SDI are requiered to confer the specific transcription. A transient transfection of HepG2 cells with various sizes of the 5'-flanking sequences, suggested the rpesence of positive and negative transcription activation region. The delination of these elements and their role in response to growth hormone is an important direction for future research.
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