Structural analysis of the supramolecular complexes involved in DNA replication
| Project/Area Number |
24770157
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| Research Category |
Grant-in-Aid for Young Scientists (B)
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| Allocation Type | Multi-year Fund |
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| Research Field |
Biophysics
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| Research Institution | National Institute of Genetics |
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Principal Investigator |
HIZUME Kohji 国立遺伝学研究所, 細胞遺伝研究系, 助教 (10378846)
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| Project Period (FY) |
2012-04-01 – 2015-03-31
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| Project Status |
Completed (Fiscal Year 2014)
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| Budget Amount *help |
¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
Fiscal Year 2014: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2013: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2012: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
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| Keywords | DNA replication / nucleosome / atomic force microscopy / DNA複製 / ヌクレオソーム / 原子間力顕微鏡 / クロマチン |
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| Outline of Final Research Achievements |
To complete correctly the replication of the genomic DNA, the initiation of DNA replication is highly regulated by the sequential association of protein complexes on replication origin.
In this study, we assembled these protein complexes on the DNA in vitro and formed relatively huge supramolecular complexes. Both molecular imaging using atomic force microscopy (AFM) and biochemical assays show that nucleosome formation on the DNA containing specific sequence of origin induces stable association of ORC on origin. We also found that nucleosome positioning around origin is rearranged by the addition of ORC.
we also performed observation of MCM-DNA complex by AFM and detected that MCM complex bound to more than two DNA strands was detected, which formed DNA-loop. This result suggests existence of the mechanism of that MCM interacts with higher-order structure of DNA such as DNA-loop rather than that MCM is simply loaded on DNA besides ORC.
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Report
(4 results)
Research Products
(9 results)
