| Budget Amount *help |
¥128,570,000 (Direct Cost: ¥98,900,000、Indirect Cost: ¥29,670,000)
Fiscal Year 2014: ¥23,920,000 (Direct Cost: ¥18,400,000、Indirect Cost: ¥5,520,000)
Fiscal Year 2013: ¥25,090,000 (Direct Cost: ¥19,300,000、Indirect Cost: ¥5,790,000)
Fiscal Year 2012: ¥26,260,000 (Direct Cost: ¥20,200,000、Indirect Cost: ¥6,060,000)
Fiscal Year 2011: ¥26,260,000 (Direct Cost: ¥20,200,000、Indirect Cost: ¥6,060,000)
Fiscal Year 2010: ¥27,040,000 (Direct Cost: ¥20,800,000、Indirect Cost: ¥6,240,000)
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| Outline of Final Research Achievements |
Structural biology has played an important role in understanding mechanism of DNA repair or replication enzymes. However, structural bases for enzyme mechanisms through chromatin remodeling are not known. Furthermore, visualization of the catalytic reaction process at the atomic level remains unclear.
In this study, we have determined the crystal structure of one subunit in an ATP-dependent chromatin remodeling factor and suggested the overall shape combined with a lot of physicochemical, biochemical and molecular biological methods. The pathway of nucleotidyl transfer reaction by human DNA polymerase eta, a key enzyme in translesion synthesis in human cells was newly visualized using time-resolved protein crystallography by freeze trapping. In sequence, the substrate and two Mg ions are aligned, the nucleophile 3'-OH is deprotonated, the sugar conformation changes and the new bond is formed. An unexpected third Mg ion arrives with the new bond and stabilizes the intermediate state.
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