| Project/Area Number |
12680668
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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 | Tohoku University |
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Principal Investigator |
KOMURA Jun-ichiro Tohoku University, Graduate School of Medicine, Research Associate, 大学院・医学系研究科, 助手 (10215410)
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| Project Period (FY) |
2000 – 2001
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥3,100,000 (Direct Cost: ¥3,100,000)
Fiscal Year 2001: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 2000: ¥1,600,000 (Direct Cost: ¥1,600,000)
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| Keywords | chromatin / nucleosome / psoralen / ionizing radiation / フットプリント |
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| Research Abstract |
1. I have developed a new genomic sequencing method for analyzing translational positioning of nucleosomes with resolution at the level of nucleotides along single-copy genes in mammalian cells, using 4,5',8-trimethylpsoralen and terminal transferase-dependent PCR. Cross-links induced by trimethylpsoralen were visualized on sequencing gels. In the regions occupied by nucleosomes, suppression of cross-linking was observed.
2. I have also developed a new method for analyzing rotational positioning of DNA on the surface of nucleosomes in living cells, utilizing X-rays and ligation-mediated PCR. DNA single-strand breaks with 5'-phosphates were visualized on sequencing gels. Repeated cutting patterns with a periodicity of 10 - 11 bases were observed in the regions occupied by rotationally positioned nucleosomes.
3. To understand the influence of the chemical modification of DNA or histone on nucleosome positioning, I have analyzed a nucleosome positioned in the promoter of the human c-FOS gene. No changes in the translational or rotational position of this nucleosome were detected upon induction of the gene, in spite of rapid acetylation and phosphorylation of histone in this region.
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