1994 Fiscal Year Final Research Report Summary
Analysis of the Regulatory Region with Unusual structural Features of the Human Thymidylate Synthase Gene
| Project/Area Number |
05671835
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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 |
Biological pharmacy
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| Research Institution | University of Shizuoka |
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Principal Investigator |
TAKEISHI Keiichi University of Shizuoka, School of Food and Nutritional Sciences, Professor, 食品栄養科学部, 教授 (90012608)
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| Co-Investigator(Kenkyū-buntansha) |
HORIE Nobuyuki University of Shizuoka, School of Food and Nutritional Sciences, Research Associ, 食品栄養科学部, 助手 (70209287)
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| Project Period (FY) |
1993 – 1994
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| Keywords | Thymidylate synthase / CAT assay / DNA element / DNA binding protein / RNA binding protein / DNA polymorphism / Differentiation / Nuclear factor |
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| Research Abstract |
In this research project, we mainly analyzed in detail regulatory elements and cellular protein factors involved in the regulation of the expression of human thymidylate synthase (TS) gene. It must have unique control features, since it is required for highly regulated expression in a cell.
First of all, we prepared many mutant DNA fragments from a specific DNA fragment derived from the region around the transcriptional initiation site of the human TS gene and analyzed promoter and cis-acting elements using the mutant DNA fragments. As the result, we identified promoter elements and several other regulatory elements in the region upstream from the major cap site. An unusual inverted repeat including a triple tandem repeat present in the region immediately upstream from an initiation codon ATG was also found to be necessary for the efficient expression of the human TS gene. In addition, the triple tandem repeat was revealed to be polymorphic with respect to the number of a repeat unit and the polymorphic DNAs were shown to differ in the expression activity of the human TS gene. Furthermore, a protein factor that bindsto an RNA fragment which corresponds to the region containing the inverted repeat was found to be present both in nuclear and cytoplasmic extracts. Finally, when human leukemia HL-60 cells differentiated into granulocytes by the retinoic acid treatment, a nuclear factor NF-TS3, that was found in our previous study and that binds to the region around the initiation codon ATG of the human TS gene was shown to increase soon after the cells started to differentiate.
We think that it is important to identify additional regulatory elements and protein factors involved in the regulation of the human TS gene expression and to reveal their roles, especially the quantitative change of the positive and negative regulatory factors in response to the alteration of the cellular environment.
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