2012 Fiscal Year Final Research Report
In-situ real-time measurement of biological informations using single molecule imaging
| Project Area | The physicochemical field for genetic activities |
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| Project/Area Number |
20114004
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| Research Category |
Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
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| Allocation Type | Single-year Grants |
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| Review Section |
Biological Sciences
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| Research Institution | Tokyo Institute of Technology |
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Principal Investigator |
TOKUNAGA Makio 東京工業大学, 大学院・生命理工学研究科, 教授 (00192659)
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| Co-Investigator(Renkei-kenkyūsha) |
SOGAWA Kumiko (SAKATA Kumiko) 東京工業大学, 大学院・生命理工学研究科, 准教授 (20291073)
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| Project Period (FY) |
2008 – 2012
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| Keywords | 1分子イメージング / 1分子計測 / 分子システム / その場計測 / 3次元イメージング / マルチカラーイメージング / HILO 照明法 / 分子定量 |
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| Research Abstract |
We have demonstrated that clear visualization of single molecules in cells enables their molecular quantification. The object of this research is to clarify structures and their changes of DNA and protein complexes that are formed transiently and locally. We examined fluorescent protein-labeled nuclear proteins, such as transcription factors, RNA polymerase II and nucleosome remodeling factors, by simultaneous multi-color single-molecule imaging using HILO microscopy. Quantification on molecular dynamics and interactions with multi-dimensional analysis and 10-nm-resolution analysis of the images enabled us to classify states of the proteins and interactions. This approach provides a tool to understand yet mystifying mechanism of regulation and expression of genetic information.
In parallel, we carried out single-molecule force measurement and molecular dynamics simulations on the same subjects. Quasi-statical unfolding of single molecules of staphylococcal nuclease and double-stranded DNA by constant-rate mechanical stretching. The mean energies to disrupt elementary structures were a few times of the thermal energy (1～3 kBT), which agrees with their stochastic feature. The results showed that entropy is a key property on the “genetic information field”.
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[Journal Article] Vesnarinone Suppresses TNF α mRNA Expression by Inhibiting Valosin-containing Protein2013
Author(s)
Hotta K, Nashimoto A, Yasumura E, Suzuki M, Azuma M, Iizumi Y, Shima D, Nabeshima R, Hiramoto M, Okada A, Sakata-Sogawa K, Tokunaga M, Ito T, Ando H, Sakamoto S, Kabe Y, Aizawa S, Imai T, Yamaguchi Y, Watanabe H, Handa H
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Journal Title
Mol Pharmacol
Volume: Vol.83
Pages: 930-938
DOI
Peer Reviewed
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[Journal Article] Vav links the T cell antigen receptor to the actin cytoskeleton and T cell activation independently of intrinsic Guanine nucleotide exchange activity2009
Author(s)
Miletic AV, Graham DB, Sakata-Sogawa K, Hiroshima M, Hamann MJ, Cemerski S, Kloeppel T, Billadeau DD, Kanagawa O, Tokunaga M, Swat W
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Journal Title
PLoS One
Volume: Vol.4
Pages: e6599
DOI
Peer Reviewed
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