| Project/Area Number |
04044133
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| Research Category |
Grant-in-Aid for international Scientific Research
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| Allocation Type | Single-year Grants |
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| Section | Joint Research |
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| Research Institution | KUMAMOTO UNIVERSITY |
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Principal Investigator |
HIRAGA Sota Kumamoto University Medical School, Professor, 医学部, 教授 (40027321)
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| Co-Investigator(Kenkyū-buntansha) |
BEGG Kenneth j. University of Edinburgh, Assistant Professor, 上級講師
DONACHIE Wil エジンバラ大学, 助教授
VINELLA Daniel University Paris VII, Instructor, ジャクモノ研究所, 助手
BOULOC Philippe University Paris VII, Instructor, ジャクモノ研究所, 助手
JAFFE Aline University Paris VII, Associate Professor, ジャクモノ研究所, 助教授
YAMANAKA Kunitoshi Kumamoto University Medical School, Instructor, 医学部, 助手 (90212290)
NIKI Hironori Kumamoto University Medical School, Lecturer, 医学部, 講師 (70208122)
OGURA Teru Kumamoto University Medical School, Associate Professor, 医学部, 助教授 (00158825)
DONACHIE William d. University of Edinburgh, Associate Professor
KENIIETH J.B エジンバラ大学, 上級講師
WILLIAM.D.DO ドナヒー エジンバラ大学, 助教授
DANIEL Vinel パリ第7大学, ジャクモノ研究所, 助手
PHILIPPE Bou パリ第7大学, ジャクモノ研究所, 助手
ALINE Jaffe パリ第7大学, ジャクモノ研究所, 助教授
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| Project Period (FY) |
1992 – 1993
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| Project Status |
Completed (Fiscal Year 1993)
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| Budget Amount *help |
¥10,200,000 (Direct Cost: ¥10,200,000)
Fiscal Year 1993: ¥5,100,000 (Direct Cost: ¥5,100,000)
Fiscal Year 1992: ¥5,100,000 (Direct Cost: ¥5,100,000)
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| Keywords | chromosome partitioning / muk gene / ftsH gene / ATPase / protease / cold shock protein / DNA binding protein / membrane protein / 大腸菌 / MukB蛋白 / ATP / GTP結合能 / DNA結合能 / FtsH蛋白 |
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| Research Abstract |
mukB mutants of Escherichia coli are defective in the correct partitioning of replicated chromosomes. This results in the appearance of normal-sized anucleate (chromosome-less) cells during cell proliferation. Based on the nucleotide sequence of the mukB gene, the MukB protein of 177 kDa was predicted to be a filamentous protein with globular domains at the ends, and also having DNA binding and nucleotide binding abilities. The purified MukB protein possesses these characteristics. MukB forms a homodimer with a rod-and-hinge structure having a pair of large, C-terminal globular domains at one end and a pair of small, N-terminal globular domains at the opposite end ; it tends to bend at a middle hinge site of the rod section. Chromatography in a DNA-cellulose column and the gel retardation assay revealed that MukB possesses DNA binding activity. Photoaffinity cross-linking experiments showed that MukB binds to ATP and GTP in the presence of Zn^<2+>. Throughout the purification steps, acyl carrier protein was co-purified with MukB.
The ftsH gene is essential for cell viability in Escherichia coli. We cloned and sequenced the wild-type ftsH gene and the temperature-sensitive ftsHl (Ts) gene. It was suggested that FtsH protein was an integral membrane protein of 70.7 kDa (644 amino acid residues) with a putative ATP-binding domain. The ftsHl(Ts) gene was found to have two base substitutions within the coding sequence corresponding to the amino acid substitutions Glu-463 by Lys and Pro-587 by Ala. Homology search revealed that an -200-amino-acid domain, including the putative ATP-binding sequence, is highly homologous (35 to 48% identical) to the domain found in members of a novel, eukaryotic family of putative ATPases, e.g., Sec18p, Pas1p, CDC48p, and TBP-1, which function in protein transport pathways, peroxisome assembly, cell division cycle, and gene expression, respectively. Possible implications of these observations are discussed.
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