Direct single-molecule observations of DNA unwinding by SV40 large tumor antigen under a negative DNA supercoil state
| Project/Area Number |
15K06946
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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 |
Molecular biology
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| Research Institution | Gunma University |
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Principal Investigator |
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| Project Period (FY) |
2015-10-21 – 2018-03-31
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| Project Status |
Completed (Fiscal Year 2017)
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| Budget Amount *help |
¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000)
Fiscal Year 2017: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2016: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2015: ¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000)
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| Keywords | DNA / DNA複製 / 高次構造 / 超らせん / SV40 / １分子観察 / １分子解析 / 複製起点 / 複製開始 / DNAポリメラーゼ / DNA合成反応 / T4 DNAリガーゼ / １分子計測 / 分子計測 / 分子操作 / 可視化技術 / 酵素反応 |
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| Outline of Final Research Achievements |
Superhelices, which are induced by the twisting and coiling of double-helical DNA in chromosomes, are thought to affect DNA metabolic processes. In this study, I report the effects of negative supercoiling on the unwinding activity of simian virus 40 large tumor antigen (SV40 TAg) at a single-molecular level. The supercoiling density of linear DNA templates was controlled using magnetic tweezers and monitored using a fluorescent microscope in a flow cell. SV40 TAg-mediated DNA unwinding under relaxed and negative supercoil states was analyzed by the direct observation of both single- and double-stranded regions of single DNA molecules. Increased negative superhelicity stimulated SV40 TAg-mediated DNA unwinding more strongly than a relaxed state; furthermore, negative superhelicity was associated with an increased probability of SV40 TAg-mediated DNA unwinding. These results suggest that negative superhelicity helps to regulate the initiation of DNA replication.
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Report
(4 results)
Research Products
(10 results)
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[Presentation] Direct single-molecule observations of DNA synthesis and digestion reactions.2015
Author(s)
Shunsuke Takahashi, Shohei Kawasaki, Hirofumi Kurita, Takeshi Mizuno, Shun-ichi Matsuura, Akira Mizuno, Masahiko Oshige, Shinji Katsura
Organizer
The 11th International Workshop on Radiation Effects on Semiconductor Devices for Space Applications (11th RASEDA) and The 7th International Conference on Advanced Micro-Device Engineering (7th AMDE)
Place of Presentation
桐生市市民文化会館（群馬県・桐生市）
Year and Date
2015-11-12
Related Report
Int'l Joint Research
