| Project/Area Number |
26440023
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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 |
Structural biochemistry
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| Research Institution | University of Yamanashi |
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Principal Investigator |
OYAMA Takuji 山梨大学, 総合研究部, 准教授 (60423133)
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| Co-Investigator(Renkei-kenkyūsha) |
ISHINO Yoshizumi 九州大学, 農学研究院, 教授 (30346837)
SHIRAI Tsuyoshi 長浜バイオ大学, バイオサイエンス学部, 教授 (00262890)
MAYANAGI Kouta 九州大学, 生体防御医学研究所, 助教 (50418571)
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| Project Period (FY) |
2014-04-01 – 2017-03-31
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| Project Status |
Completed (Fiscal Year 2016)
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| Budget Amount *help |
¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000)
Fiscal Year 2016: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2015: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2014: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
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| Keywords | DNA複製 / タンパク質DNA複合体 / 結晶構造解析 |
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| Outline of Final Research Achievements |
To understand the detailed loading mechanism of the clamp PCNA onto DNA by the clamp-loader RFC in an ATP-dependent manner, we aimed (1) preparation of selenomethionine-labeled (SeMet) PCNA, (2) high resolution structure determination of RFC, and (3) crystallization and structure determination of the RFC-PCNA-DNA ternary complex. We determined the crystal structure of SeMet PCNA at 2.5 angstrom resolution. We established a novel preparation method of the highly purified recombinant RFC, but did not succeed in the high-resolution structure determination of the RFC. We performed crystallization screening of the RFC-PCNA-DNA ternary complex in a wide range of conditions, but obtained no crystals during this research project.
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