1997 Fiscal Year Final Research Report Summary
Molecular biological studies on the induction mechanism of erythropoietin by hypoxic signal
| Project/Area Number |
07457554
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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 |
Human genetics
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| Research Institution | Tokyo Medical and Dental University |
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Principal Investigator |
YASUKOCHI Yukio Tokyo Medical and Dental University, Medical Research Institute, Dept. of Molecular Genetics, Professor, 難治疾患研究所, 教授 (60037398)
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| Co-Investigator(Kenkyū-buntansha) |
TSUCHIYA Terumasa Tokyo Medical and Dental University, Medical Research Institute, Dept. of Molecu, 難治疾患研究所, 助手 (20242109)
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| Project Period (FY) |
1995 – 1997
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| Keywords | Oxygen senser / hypoxia / HNF-4 / mRNA differential display / erythropoietin / GATA motif |
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| Research Abstract |
1) studies on oxygen senser
First we investigated a possible existance of the kinase type of oxygen senser. By exposing Hep 3B cells to cobalt in place of hypoxia, we cloned one cDNA which was subjected to specific phosphrylation. Second we pursued the transcription type of oxygen senser and found HNF-4 be a candidate for it.
2) Oxygen and metal responsive gene superfamily
We obtained one and five novel clones by differential screening and mRNA differential display methods, respectively, which were up-regulated in hypoxia or exposure to cobalt. The former contained the LXXLL motif which indicate a possible transcriptional coactivator. Also the recombinant protein produced in E.coli could interact with various transcription factors including the basal transcription factors.
3) Control mechanism of erythropoietin gene expression
The GATA motif in the -30 region of the transcription initiation site of the gene is highly concerved among species while the consensus motif in the -30 region is TATA in many genes. When the GATA motif was mutated to TATA,the transcriptional activity of the mutant was enhanced even in normoxia. Binding assay showed that TATA-binding protein (TBP) could weakly bind to the wild-type GATA motif whereas TBP bound to the mutant TATA motif with high affinity.
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