| Project/Area Number |
07557153
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 試験 |
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| Research Field |
Biological pharmacy
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| Research Institution | Tohoku University |
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Principal Investigator |
ENOMOTO Takemi Tohoku university, Faculty of Pharmaceutical Sciences, Professor, 薬学部, 教授 (80107383)
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| Co-Investigator(Kenkyū-buntansha) |
OKUYAMA Akira Banyu Pharmaceutical Co. , LTD Molecular Biology Research Laboratories, Director, つくば研究所, ディレクター
SEKI Masayuki Tohoku university, Faculty of Pharmaceutical Sciences, Assistant professor, 薬学部, 助手 (70202140)
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| Project Period (FY) |
1995 – 1996
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| Project Status |
Completed (Fiscal Year 1996)
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| Budget Amount *help |
¥13,600,000 (Direct Cost: ¥13,600,000)
Fiscal Year 1996: ¥5,600,000 (Direct Cost: ¥5,600,000)
Fiscal Year 1995: ¥8,000,000 (Direct Cost: ¥8,000,000)
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| Keywords | DNA replication / DNA helicase / DNA-dependent ATPase / inhibitor / screening / anti-cancer drug / mammalian cells |
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| Research Abstract |
The aim of this project is to provide the basis for developing new anti-cancer drugs by identifying DNA helicase involved in DNA replication in mammalian cells and by developing screening systems for inhibitors of the helicase. We have succeeded to identify the DNA helicase involved in DNA replication in mammalian cells by obtaining following results.
1. In tsFT848 cells that are temperature-sensitive mutants for DNA replication, the decrease in the level of DNA synthesis correlated with the decrease in the level of DNA-dependent ATP ase activity of DNA helicase B.
2. DNA helicase B from tsFT848 cells was more heat-sensitive than that from wild-type cells.
3. One Missense mutation from Phe to Cys occurred in the DNA helicase B of mutant cells at the amino acid position, 724.
4.DNA helicase B stimulated DNA primase activity.
5.DNA helicase B functioned as a helicase in a cell-free DNA replication system.
6. DNA helicase B was localized at the replication foci.
As to the development of screening systems, we developed a system measuring DNA-dependent ATPase activity instead of DNA helicase activity with a platereader, which was able to deal with many samples at once. To supply the target helicase to the screening system, we have tried to express DNA helicase B in Escherichia coli and yeast but not succeeded yet. We are now trying to obtain large amount of DNA helicase B by using the baculovirus system.
Although we have not succeeded to find inhibitors for DNA helicase B,we think that we could provide the basis for developing new anti-cancer drugs by identifying the target and developing the screening system.
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