Analysis of Structure and Function of Human Centromere/kindochore Chromatin
| Project/Area Number |
15370076
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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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 | Nagoya University |
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Principal Investigator |
YODA Kinya Nagoya University, Bioscience and Biotechnology Center, Associate Professor, 生物機能開発利用研究センター, 助教授 (30126916)
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| Co-Investigator(Kenkyū-buntansha) |
OBUSE Chikasihi Kyoto University, Graduate School of Biostudies, Associate Professor, 大学院・生命科学研究科, 助教授 (00273855)
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| Project Period (FY) |
2003 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥12,300,000 (Direct Cost: ¥12,300,000)
Fiscal Year 2004: ¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2003: ¥8,800,000 (Direct Cost: ¥8,800,000)
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| Keywords | human centromere chromatin / anti-CENP-A monoclonal antibody / I-CEN complex / proteomics analysis / siRNA / MgcRacGAP / ASR2A / Rsf-1 / SNF2H remodeling complex / SNF2Hリモデリング複合体 / N-ChIP / 機能解析 / CENP-A / B / C 複合体 / I型αサテライトDNA / クロマチンリモデリング因子 / ポリコーム複合体 |
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| Research Abstract |
Our Group has been studying about structure and function of the centromere region of human chromosomes in the viewpoint of DNA/protein complex formed by interaction between DNA and proteins. We have elaborated to purify a centromere specific histone H3 variant, CENP-A, and established the method for reconstruction of the nucleosome containing CENP-A instead of histone H3 (CENP-A nucleosome)(ref.6). We have produced highly specific anti-CENP-A monoclonal antibody and this antibody has been supplied to lots of investigators inside and/or outside of this country (ref.1,11-13). We have shown that this antibody was excellent in high specificity not only for immuno-stain of the cells but also for chromatin immuno-precipitation (ref.4). We have developed the method to mildly solubilize the chromatin and isolate the centromere chromatin from interphase nuclei using anti-CENP-A antibody (Native Chromatin Immuno-Precipitation;N-ChIP)(ref.9). The isolated chromatin complex contained all the reported centromere structural proteins, CENP-A,CENP-B,CENP-C,CENP-H,CENP-l/hMis6 and hMis12. Therefore, we call this complex as Interphase CENtromere complex(I-CEN complex). The components of the I-CEN complex have been comprehensively subjected to LC/MS/MS analyses and 40 proteins were detected (ref.9). In collaboration with Dr.Naohito Nozaki, Kanagawa Dental College, we tried to produce the antibodies against 20 proteins using chemically synthesized amino-and/or carboxyl-terminal peptides as antigens. We've got 10 kinds of antibodies and 5 of them are now under analysis ; antibodies against MgcRacGAP,hMKLP-1,ASR2A,Rsf1(XAP8) and SNF2H. To analyze the physiological function of each protein we are now trying RNA-interference-mediated depletion of the proteins. By siRNA transfection we reveal that ASR2A is essential for interaction between kinetochore and microtubule (paper in preparation).
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Report
(3 results)
Research Products
(37 results)
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[Journal Article] Inhibitors of histone deacetylases alter kinetochore assembly by disrupting pericentromeric heterochromatin.2005
Author(s)
Robbins A.R., Jablonski, S.A., Yen, T.J., Yoda, K., Robey, R., Bates, S.E., Sackett, D.L.
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Journal Title
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Centromere protein H is upregulated in primary human colorectal cancer and its overexpression induces aneuploidy.2005
Author(s)
Tomonaga, T., Matsushita, K., Ishibashi, M., Nezu, M., Shimada, H., Ochiai, T., Yoda.K., Nomura, F.
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Journal Title
Cancer Res. 65(11)
Pages: 4683-4689
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Expression and purification of recombinant human histones.2004
Author(s)
Tanaka, Y., Tawaramoto-Sasanuma, M., Kawaguchi, S., Ohta, T., Yoda, Kurumizaka, H., Yokoyama, S.
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Description
「研究成果報告書概要(欧文)」より
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