| Project/Area Number |
13043029
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| Research Category |
Grant-in-Aid for Scientific Research on Priority Areas
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| Allocation Type | Single-year Grants |
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| Review Section |
Biological Sciences
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| Research Institution | Osaka University |
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Principal Investigator |
TAKISAWA Haruhiko Osaka University, Graduate School of Science, Department of Biological Sciences, Professor, 大学院理学研究科, 教授 (60154944)
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| Co-Investigator(Kenkyū-buntansha) |
SHIRAHIGE Katsuhiko Tokyo Institute of Technology, Gene Research Center, Associate Professor, バイオ研究基盤支援総合センター, 助教授 (90273854)
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| Project Period (FY) |
2001 – 2005
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| Project Status |
Completed (Fiscal Year 2005)
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| Budget Amount *help |
¥75,200,000 (Direct Cost: ¥75,200,000)
Fiscal Year 2005: ¥16,000,000 (Direct Cost: ¥16,000,000)
Fiscal Year 2004: ¥15,600,000 (Direct Cost: ¥15,600,000)
Fiscal Year 2003: ¥14,400,000 (Direct Cost: ¥14,400,000)
Fiscal Year 2002: ¥15,000,000 (Direct Cost: ¥15,000,000)
Fiscal Year 2001: ¥14,200,000 (Direct Cost: ¥14,200,000)
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| Keywords | DNA replication / Initiation complex / GINS / rep liso me / ChIP-chip method / Replication checkpoint / DNA複製因子 / 複製ライセンス化 / 複製チェックポイント / RecQ4 / Sld2 / Tof1 / Tim1 / Csm3 / Tipin / 複製フォーク / Mrc1 / geminin / Dpb11 / Cut5 / TopBP1 / DNA複製制御 / S期CDK / アフリカツメガエル / 無細胞複製系 / MCM / Cdc45 / MCM蛋白質 / DNAヘリカーゼ / Mcmタンパク質 |
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| Research Abstract |
UsingXenopuseggextracts and budding yeast,we investigatedtheregulationof initiation andelongation stages of DNA replication. We have clarified conserved and diverged regulation mechanisms for DNA replication.
We have identified a novel and highly conserved replication protein complex, GINS, as an essential component required for the assembly of initiation machinery of DNA replication. We have also identified conserved components of replisomes as GINS, Cdc45, Tofl/Timl, Csm3/Tipin and Mrcl/claspin. We have also identified Xenopus homolog of yeast Cut5/Dpb 11 and Sld2/Drc 1 as Xenopus CutS and RecQ4. These proteins show diverged structure and function compared with yeast proteins, suggesting that these proteins gain some additional functions during evolution of metazoans.
We developed a high-density oligonucleotide array that was able to analyze yeast chromosome VI at a 300-base pair (bp) resolution and introduced ChIP-chip (Chromatin immuno-precipitation combined with DNA chip) as a very powerful tool to visualize the dynamic changes of replication proteins and progression of DNA replication. This technology opens up the way to understand how local protein-protein or protein-DNA interactions lead to the dynamic changes of chromosome structure and clarified 1) how DNA replication and cell cycle checkpoint are coordinated, and 2) how DNA transcription is coordinated with sister chromatid cohesion.
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