| Project/Area Number |
11694277
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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 | 一般 |
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| Research Field |
Molecular biology
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| Research Institution | Osaka University |
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Principal Investigator |
SUGINO Akio Research Institute for Microbial Diseases, Osaka University, Professor, 微生物病研究所, 教授 (90231737)
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| Co-Investigator(Kenkyū-buntansha) |
HIRAGA Shin-ichiro Research Institute for Microbial Diseases, Osaka University, Assistant Professor, 微生物病研究所, 助手 (90322172)
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| Project Period (FY) |
1999 – 2001
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥19,110,000 (Direct Cost: ¥17,400,000、Indirect Cost: ¥1,710,000)
Fiscal Year 2001: ¥7,410,000 (Direct Cost: ¥5,700,000、Indirect Cost: ¥1,710,000)
Fiscal Year 2000: ¥5,600,000 (Direct Cost: ¥5,600,000)
Fiscal Year 1999: ¥6,100,000 (Direct Cost: ¥6,100,000)
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| Keywords | Chromosomal DNA replication / Mcm2-7 protein complex / Mcm10 protein / S-phase checkpoint / Rad53p / Cdc7 / Dbf4 protein kinase complex / Protein kinase / Xenopus egg in vitro replication system / MCM10蛋白 / Cdc7p / Dbf4p複合体 / in vitro 複製系 / Xenopus egg / DNAポリメラーゼ / Sー期チェックポイント |
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| Research Abstract |
Saccharomyces cerevisiae Cdc7/Dbf4 protein kinase, which regulates the initiation of chromosomal DNA replication, was expressed and purified from insect cells. It was found that the purified Cdc:7/Dbf4 phosphorylates Mcm2, which is one of Mcm2-7 complex. Furthermore, its phosphorylation sites were determined. Xenopus homologs of S. cerevisiae Cdc7/Dbf4 protein Kinase and Mcm10 were identified and their corresponding genes were cloned. it was found that Xenopus Cdc7/Dbf4.protein kinase and Mcm10 are required for the initiation of chromosomal DNA replication as in S. cerevisiae.
S. cerevisiae Rad53p, which involved in both DNA-damage-and S-phase checkpoint regulation, was purified and one of the substrates was identified to be Dbf4/Cdc7 protein kinase. It was found that Rad53p directly intercts with Cdc7/Dbf4 and phosphorylates Dbf4. Phosphorylation of Cdc7/Dbf4 complex with Rad53p inactivates Cdc7/Dbf4 protein kinase activity.
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