Structural study of the higher order chromatin structure formed by HP1BP3
| Project/Area Number |
15H06692
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| Research Category |
Grant-in-Aid for Research Activity Start-up
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| Allocation Type | Single-year Grants |
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| Research Field |
Structural biochemistry
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| Research Institution | Waseda University |
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Principal Investigator |
Koyama Masako 早稲田大学, 理工学術院, 次席研究員(研究院助教) (40755097)
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| Project Period (FY) |
2015-08-28 – 2017-03-31
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| Project Status |
Completed (Fiscal Year 2016)
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| Budget Amount *help |
¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000)
Fiscal Year 2016: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2015: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
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| Keywords | クロマチン / ヌクレオソーム / 分裂酵母 / リンカーヒストン / ヒストン / HP1BP3 / X線結晶解析 |
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| Outline of Final Research Achievements |
We have aimed to determine the structure of the fundamental unit of higher order chromatin formed by human HP1BP3, the recently identified linker histone in human. However, it was difficult to purify the HP1BP3 protein at high yield necessary for the structural studies. Therefore, we selected the functional homolog of HP1BP3 in S. pombe. The protein was expressed as a soluble recombinant protein in E. coli, and purified in high quality. Furthermore, we established the method to reconstitute S. pombe nucleosomes in vitro. We purified the four canonical core histones in S. pombe (H2A, H2B, H3, and H4, which share 79%, 68%, 92%, and 91% amino acid sequence homology with their human counterparts). Using S. pombe histones and a 146-base-pair DNA, we successfully reconstituted nucleosomes in vitro. We confirmed that the HP1BP3 homolog in S. pombe bind the nucleosome.
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Report
(3 results)
Research Products
(5 results)
