| Project/Area Number |
17570148
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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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 | The Institute of Physical and Chemical Research |
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Principal Investigator |
IZUMI Masako The Institute of Physical and Chemical Research, Radiation Biology Team, 専任研究員 (00280719)
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| Project Period (FY) |
2005 – 2007
|
| Project Status |
Completed (Fiscal Year 2007)
|
| Budget Amount *help |
¥3,200,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥300,000)
Fiscal Year 2007: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2006: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 2005: ¥1,000,000 (Direct Cost: ¥1,000,000)
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| Keywords | Mcm 10 / DNA replication / cell cycle |
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| Research Abstract |
Mcm10 was first identified in S. cerevisiae as an essential protein for chromosomal DNA replication. Recent studies have shown that Mcm10 has multiple functions both in the initiation of DNA replication and the replication elongation, although the precise roles of Mcm10 are still unknown. Recently we have hind that GFP-tagged human Mcm10 fanned discrete foci in the nuclei, and the distribution patterns of GFP-Mcm10 foci preceded changes in replication of patterns by 30〜60 min. These results suggest that human Mcm10 is recruited to the replication sites 30-60 min before they replicate. In this study, we have mapped a domain required for its foci formation by using the truncated derivatives of the fill-length 874-amino acid human Mcm10. The retention of Mcm10 onto chromatin required the domain containing amino acids 473-693, which was overlapped with the domain interacting with Mcm2-7. Depletion of Mcm10 by siRNA slowed down the S phase progression and interfered the chromatin loading of RPA and PCNA, although Cdc45 and DNA polymerases were loaded onto chromatin. The delay of S phase progression was caused by the inhibition of new origin firing, not by the inhibition of established replication fork. These results suggest that human Mcm10 is loaded onto the chromatin through the interaction with Mcm2-7 and involved in the activation of pre-replication complex after Cdc45 and DNA polymerases are loaded onto chromatin
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