| Project/Area Number |
15590362
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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 |
Experimental pathology
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| Research Institution | Institute for Developmental Research, Aichi Human Service Center |
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Principal Investigator |
NAKAYAMA Atsuo Institute for Developmental Research, Aichi Human Service Center, Dept Embryology, Department Chief, 発生障害学部, 部長 (50227964)
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| Project Period (FY) |
2003 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥2,900,000 (Direct Cost: ¥2,900,000)
Fiscal Year 2004: ¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 2003: ¥1,300,000 (Direct Cost: ¥1,300,000)
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| Keywords | RFX1 / transcriptional regulation / p120^<E4F> / MAP1A promoter / PCNA promoter / Adenovirus E1A / reporter assay / chromatin immunoprecititation / RFX1 / E4F1 / 転写補助因子 |
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| Research Abstract |
To elucidate the complex mechanisms, in which REX transaction factors activate and inactivate the transcription depending on the gene and cell types, we searched factors interacting with REX by yeast two hybrid.
We obtained about 300 positive clones in the yeast system, and 180 clones of them were sequenced. Eighteen clones were further investigated by biochemical methods whether they bind to RFX, and a cellar transcription factor, p120^<E4F> was identified as a protein interacting with RFX1. Co-immunoprecipitation using series of deletion constructs indicated that the cluster of Zinc fingers in the middle portion of p120^<E4F>, and a region containing the DNA binding domain of RFX1 are necessary for the interaction. Reporter assay indicated that p120^<E4F> counteracts to the repressive effects by RFX1 on the MAP1A promoter, but p120^<E4F> itself has no effect on the MAP1A promoter. Thus the effect of p120^<E4F> depends on the interaction between RFX1 and p120^<E4F>. Chromatin DP showed that overexpression of p120^<E4F> did not clearly affect the binding of RFX1 to the target gene, suggesting that the way of counteraction to repression, is not by obstructing DNA-binding of RFX1. In addition to the MAP1A promoter, p120^<E4F> has considerable effects on the PCNA promoter, which is under control of RFX1.
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