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1993 Fiscal Year Final Research Report Summary

Prokaryotic force-generating enzyme and protein transport.

Research Project

Project/Area Number 04044133
Research Category

Grant-in-Aid for international Scientific Research

Allocation TypeSingle-year Grants
SectionJoint Research
Research InstitutionKUMAMOTO UNIVERSITY

Principal Investigator

HIRAGA Sota  Kumamoto University Medical School, Professor, 医学部, 教授 (40027321)

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
Project Period (FY) 1992 – 1993
Keywordschromosome partitioning / muk gene / ftsH gene / ATPase / protease / cold shock protein / DNA binding protein / membrane protein
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.

  • Research Products

    (24 results)

All Other

All Publications (24 results)

  • [Publications] Feng J.: "New killing system controlled by two genes located immediately upstream of the mukB gene in E.coli." Mol.Gen.Genet.(in press). (1994)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yamanaka K.: "Characterization of translucent segments observed in an smbA mutant of Escherichia coli." FEMS Microbiol.Lett.(in press). (1994)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yamanaka K.: "Multicopy suppressors,mssA and mssB,of an smbA mutation of Escherichia coli." Mol.Gen.Genet.(in press). (1994)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Akiyama Y.: "Involvement of FtsH in protein assembly into and through the membrane.1.Mutation that reduce retention efficiency of a cytoplasmic reporter." J.Biol.Chem.(in press). (1994)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hiraga S.: "E.coli chromosome partition." Current Opinion in Genetics & Development. 5. 789-801 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Herman C.: "Cell growth and I phage development controlled by the same essential Escherichia coli gene,ftsH/hflB." Proc.Natl.Acad.Sci.USA. 90. 10861-10865 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Tomoyasu T.: "The Escherichia coli FtsH protein is a prokaryotic member of a protein family of putative ATPases involved in membrane functions,cell cycle control and gene expression." J.Bacteriol.175. 1344-1351 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Tomoyasu T.: "Topology and subcellular localization of FtsH protein in Escherichia coli." J.Bacteriol.175. 1352-1357 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Niki H.: "E.coli MukB protein involved in chromosome partition forms a homodimer with a rod-and-hinge structure having DNA binding and ATP/GTP binding activities." EMBO J.11. 5101-5109 (1992)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yamanaka K.: "Identification and characterization of the smbA gene,a suppressor of the mukB null mutant in Escherichia coli." J.Bacteriol.174. 7517-7526 (1992)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Begg K.J.: "Escherichia coli mutant Y16 is a double mutant carrying thermosensitive ftsH and ftsl mutations." J.Bacteriol.174. 2416-2417 (1992)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Imamura R.: "Characterization of high molecular weights of complexes and polymers of cytoplasmic proteins in Escherichia coli." Res.Microbiol.143. 743-753 (1992)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Feng J.: "New killing system controlled by two genes located immediately upstream of the mukB gene in E. coli." Mol.Gen.Genet.(in press). (1994)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yamanaka K.: "Characterization of translucent segments observed in an smbA mutant of Escherichia coli." FEMS Microbiol.Lett.(in press). (1994)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yamanaka K.: "Multicopy suppressors, mssA and mssB, of an smbA mutation of Escherichia coli." Mol.Gen.Genet.(in press). (1994)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Akiyama Y.: "Involvement of FtsH in protein assembly into and through the membrane. 1. Mutation that reduces retention efficiency of a cytoplasmic reporter." J. Biol.Chem.(in press). (1994)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Hiraga S.: "E.coli chromosome partition." Current Opinion in Genetics & Development. 5. 789-801 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Herman C.: "Cell growth and I phage development controlled by the same essential Escherichia coli gene, ftsH/hflB." Proc. Natl.Acad.Sci.USA. 90. 10861-10865 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Tomoyasu T.: "The Escherichia coli FtsH protein is a prokaryotic member of a protein family of putative ATPases involved in membrane functions, cell cycle control and gene expression." J. Bacteriol.175. 1344-1351 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Tomoyasu T.: "Topology and subcellular localization of FtsH protein in Escherichia coli." J. Bacteriol.175. 1352-1357 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Niki H.: "E.coli MukB protein involved in chromosome partition forms a homodimer with a rod-and-hinge structure having DNA binding and ATP/GTP binding activities." EMBO J.11. 5101-5109 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yamanaka K.: "Identification and characterization of the smbA gene, a suppressor of the mukB null mutant in Escherichia coli." J. Bacteriol.174. 7517-7526 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Begg K.J.: "Escherichia coli mutant Y16 is a double mutant carrying thermosensitive ftsH and fts1 mutations." J. Bacteriol.174. 2416-2417 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Imamura R.: "Characterization of high molecular weights of complexes and polymers of cytoplasmic proteins in Escherichia coli." Res. Microbiol.143. 743-753 (1992)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 1995-02-07  

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