|Budget Amount *help
¥137,280,000 (Direct Cost: ¥105,600,000、Indirect Cost: ¥31,680,000)
Fiscal Year 2012: ¥19,890,000 (Direct Cost: ¥15,300,000、Indirect Cost: ¥4,590,000)
Fiscal Year 2011: ¥19,890,000 (Direct Cost: ¥15,300,000、Indirect Cost: ¥4,590,000)
Fiscal Year 2010: ¥19,890,000 (Direct Cost: ¥15,300,000、Indirect Cost: ¥4,590,000)
Fiscal Year 2009: ¥22,100,000 (Direct Cost: ¥17,000,000、Indirect Cost: ¥5,100,000)
Fiscal Year 2008: ¥55,510,000 (Direct Cost: ¥42,700,000、Indirect Cost: ¥12,810,000)
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”.