Functional analyses and development of inhibitors based on higher-order structure of translesion DNA polymerase eta
| Project/Area Number |
15H04646
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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 |
Biological pharmacy
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| Research Institution | Gakushuin University |
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Principal Investigator |
HANAOKA Fumio 学習院大学, 理学部, 研究員 (50012670)
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| Project Period (FY) |
2015-04-01 – 2018-03-31
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| Project Status |
Completed (Fiscal Year 2018)
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| Budget Amount *help |
¥16,900,000 (Direct Cost: ¥13,000,000、Indirect Cost: ¥3,900,000)
Fiscal Year 2017: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2016: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000)
Fiscal Year 2015: ¥8,710,000 (Direct Cost: ¥6,700,000、Indirect Cost: ¥2,010,000)
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| Keywords | DNAポリメラーゼ / 損傷乗り越え合成 / DNA損傷 / 紫外線感受性 / 突然変異 / 阻害剤 / スクリーニング / 遺伝子 / 癌 / CPD |
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| Outline of Final Research Achievements |
In order to identify the important amino acid residues for the template DNA binding of human DNA polymerase eta (Pol eta), we generated several point mutants of Pol eta based on the co-crystal structure of Pol eta-UV-damaged DNA complex. We analyzed these purified Pol eta biochemically as well as UV sensitivity and mutation frequency of 6-thioguanine resistance.
We also conducted screening of Pol eta inhibitors using chemical arrays of natural low molecular-weight compounds by Pol eta-binding at first and then examined inhibitory activities of these compounds using primer-extension reaction. We successfully obtained several Pol eta inhibitor candidates and some of them are plausibly specific for Pol eta.
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| Academic Significance and Societal Importance of the Research Achievements |
ヒトPolηと紫外線損傷DNAとの共結晶構造に基づいて、鋳型DNAとの相互作用に重要と考えられるアミノ酸を置換した点変異体タンパク質の解析から、鋳型DNAとの相互作用が弱い変異体は全て野生型と比較してDNA鎖の伸長反応が進みにくい結果となった。このことは結晶構造解析によるタンパク質の機能予測が非常に的確であることを示している。
一方、天然物由来の低分子化合物を固定化した化合物アレイを用い、ヒトPolηに対して阻害効果を示す化合物のスクリーニングを行い、Polηに特異的な阻害剤の候補を選択した。これらは代表的な制がん剤であるシスプラチンとの併用により、制がん効果の増強に効果的な可能性がある。
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Report
(4 results)
Research Products
(34 results)
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[Journal Article] Excision of translesion synthesis errors orchestrates responses to helix-distorting DNA lesions2015
Author(s)
Anastasia Tsaalbi-Shtylik, Cristina Ferrás, Bea Pauw, Giel Hendriks, Piya Temviriyanukul, Leone Carlée, Fabienne Calléja, Sandrine van Hees, Jun-Ichi Akagi, Shigenori Iwai, Fumio Hanaoka, Jacob G. Jansen, and Niels de Wind
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Journal Title
The Journal of Cell Biology
Volume: 209
Issue: 1
Pages: 33-46
DOI
Related Report
Peer Reviewed / Open Access
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[Journal Article] Structural basis of pyrimidine-pyrimidone (6-4) photoproduct recognition by UV-DDB in the nucleosome.2015
Author(s)
Osakabe, A., Tachiwana, H., Kagawa, W., Horikoshi, N., Matsumoto, S., Hasegawa, M., Matsumoto, N., Toga, T., Yamamoto, J., Hanaoka, F., Thoma, N.H., Sugasawa, K., Iwai, S., Kurumizaka, H.
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Journal Title
Scientific Reports
Volume: 5
Issue: 1
Pages: 16330-16330
DOI
Related Report
Peer Reviewed / Open Access / Int'l Joint Research
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