Construction of a DNA double-strand break model and its application to studies of DNA repair and low dose irradiation effects
| Project/Area Number |
25340044
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Risk sciences of radiation and chemicals
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| Research Institution | National Institute of Health Sciences |
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Principal Investigator |
Honma Masamitsu 国立医薬品食品衛生研究所, 変異遺伝部, 部長 (30250179)
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| Research Collaborator |
Yasui Manabu
Suzuki Tetsuya
Sassa Akira
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| Project Period (FY) |
2013-04-01 – 2019-03-31
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| Project Status |
Completed (Fiscal Year 2018)
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| Budget Amount *help |
¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000)
Fiscal Year 2017: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2016: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2015: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2014: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2013: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
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| Keywords | DNA修復 / 放射線防護 / 低線量モデル / 発がん / 遺伝的不安定性 / 相同組み換え / BLMヘリケース / ゲノム編集技術 / DNA 修復 |
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| Outline of Final Research Achievements |
As a model for low dose radiation, I developed a system that generates only one DNA double-strand breaks (DSB)DSB in the human cell genome using a restriction enzyme I-SceI site. This system revealed that a DSB mainly causes small deletions by non-homologous end joining (NHEJ). Involvement of homologous recombination (HR) was less than 10% of the whole. In addition, in order to elucidate the mechanism of genome integrity, I focused on recombination repair between chromosomes. I constructed BLM knocked out (KO) cells using genome editing technology in the system. In BLM-KO cells, the enhancement of spontaneous HR frequency, cross-type gene conversion, non-cross-type long gene conversion, and deletion were frequently observed. BLM was considered to be involved in the maintenance of Holliday structure, and the defect destabilizes Holliday structure, leading to the deletion and recombination of the genome in a large area.
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| Academic Significance and Societal Importance of the Research Achievements |
放射線はゲノムに対して最も重篤であるDNA二本鎖切断（DSB）を誘発し、それが、がんの主たる要因と考えられているが、放射線はそれ以外の損傷（酸化ストレス、一本鎖切断等）も引き起こすため、DSBの修復機構や、がん化への関与の研究は非常に困難である。本研究では制限酵素に注目し、たった一つのDSBをヒト細胞のゲノム中に発生させる究極の低線量モデルを世界で始めて構築した。これによりDSBには閾値が存在せず、高い確率で突然変異を誘発することを明らかにし、放射線のリスク評価の考えを前進させた。
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Report
(7 results)
Research Products
(25 results)
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[Journal Article] in vivo evidence that phenylalanine 171 acts as a molecular brake for translesion DNA synthesis across benzo[a]pyrene DNA adducts by human DNA polymerase2014
Author(s)
A. Sassa, T. Suzuki, Y. Kanemaru, N. Niimi, H. Fujimoto, A. Katafuchi, P. Gruz, M. Yasui, R.C. Gupta, F. Johnson, T. Ohta, M. Honma, N. Adachi and T. Nohmi
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Journal Title
DNA Repair
Volume: 15
Pages: 21-28
DOI
Related Report
Peer Reviewed / Open Access
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[Presentation] Tracing the fates of site-specifically introduced DNA adducts in the human genome.2014
Author(s)
Honma, M., Yasui, M., Kanemaru, Y., Kamoshita, N., Suzuki, T., Arakawa, T.
Organizer
4回アジア環境変異原学会
Place of Presentation
インド、コルカタ
Year and Date
2014-12-10 – 2014-12-12
Related Report
Invited
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[Presentation] Demonstration of non-threshold of 8-oxoG inducing genotoxicity by targeted mutagenesis.2014
Author(s)
Honma, M., Yasui, M., Kanemaru, Y., Kamoshita, N., Suzuki, T., Arakawa, T.
Organizer
第43回ヨーロッパ環境変異原学会
Place of Presentation
英国、ランカスター
Year and Date
2014-07-07 – 2014-07-09
Related Report
