| Project/Area Number |
13558090
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 展開研究 |
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| Research Field |
Molecular biology
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| Research Institution | Kyushu University |
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Principal Investigator |
KATAYAMA Tsutomu Kyushu University, Faculty of Pharmaceutical Sciences, Prof., 大学院・薬学研究院, 教授 (70264059)
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| Co-Investigator(Kenkyū-buntansha) |
KURUMIZAKA Hitoshi Waseda Univ., Faculty of Engineering and Science, Assoc.Prof., 大学院・理工学部, 助教授 (80300870)
ASO Mariko Kyushu University, Faculty of Pharmaceutical Sciences, Assis.Prof., 大学院・薬学研究院, 助手 (30201891)
UEDA Tadashi Kyushu University, Faculty of Pharmaceutical Sciences, Prof., 大学院・薬学研究院, 教授 (90184928)
ENDO Atsushi Dai-chi Pharmaceuticals Co., Researcher, 創薬第一研究所, 副主任研究員
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| Project Period (FY) |
2001 – 2003
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| Project Status |
Completed (Fiscal Year 2003)
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| Budget Amount *help |
¥11,200,000 (Direct Cost: ¥11,200,000)
Fiscal Year 2003: ¥2,700,000 (Direct Cost: ¥2,700,000)
Fiscal Year 2002: ¥4,100,000 (Direct Cost: ¥4,100,000)
Fiscal Year 2001: ¥4,400,000 (Direct Cost: ¥4,400,000)
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| Keywords | DNA replication / Cell cycle / DnaA protein / E.coli / DNA polymerase / AAAt motif / ATP / Antibiotics / DNA結合 / NMR / X線結晶解析 / AAA+ファミリー / 染色体複製制御 |
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| Research Abstract |
To establish rational process for developing novel antibiotics that targets DnaA protein, we analyzed structure-function relationship of the DNA-binding domain of DnaA, DnaA domain IV(DAD-IV). First, we overproduced and purified a truncated DnaA protein that contained only the DAD-IV. We found that DAD-IV sustains specific affinity for DNA and secondary structure by gel-retardation assay and CD analysis. Then we determined the secondary structure in solution of DAD-IV using NMR. Determined structure suggested that DAD-IV contained a three helices-bundled type helix-turn-helix motif. In addition, NMR analysis suggested that interaction sites of DAD-IV with DNA. Finally, we determined crystal structure of a complex of DAD-IV with DNA using X-ray diffraction analysis. Determined structure by this well coincided with the data obtained by NMR and by other biochemical and genetic analysis. Candidates of antibiotics were searched by the method of virtual screening, based on the structure of DAD-IV/DNA complex.
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