| Budget Amount *help |
¥18,200,000 (Direct Cost: ¥14,000,000、Indirect Cost: ¥4,200,000)
Fiscal Year 2015: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
Fiscal Year 2014: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
Fiscal Year 2013: ¥9,620,000 (Direct Cost: ¥7,400,000、Indirect Cost: ¥2,220,000)
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| Outline of Final Research Achievements |
Replicated sister chromatids are faithfully segregated into two daughter cells. Kinetochore is built at the centromeric region of the chromosome and is involved in the chromosome segregation. CENP-TWSX complex is a conserved kinetochore component and plays an important role. It contains histone fold and binds to DNA but its precise mechanism remains elusive. To elucidate the structure and function of the CENP-TWSX complex, we have undertaken biochemical and structural analyses. Using isothermal titration calorimetry, we obtained dissociation constant of the CENP-TW and CENP-SX interaction. Next, we measured interaction between DNA and CENP-TW or CENP-SX. Using gel filtration, we acquired the stoichiometry of the complex. Dissociation constants were measured by the fluorescent anisotrophy method. Furthermore, we obtained crystal structure of the FANC-M-CENP-SX complex. Together, these analyses revealed detailed molecular mechanism of the CENP-TWSX complex.
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