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Analyses of Clustered Inversion Region in Incl Plasmids.

Research Project

Project/Area Number 01540533
Research Category

Grant-in-Aid for General Scientific Research (C)

Allocation TypeSingle-year Grants
Research Field 遺伝学
Research InstitutionTokyo Metropolitan University

Principal Investigator

KOMANO Teruya  Tokyo Metropolitan University, Dept. of Biology, Associate Professor., 理学部, 助教授 (00087131)

Co-Investigator(Kenkyū-buntansha) NISIOKA Taizo  Tokyo Metropolitan University, Dept. of Biology, Associate Professor., 理学部, 助教授 (50087053)
Project Period (FY) 1989 – 1990
Project Status Completed (Fiscal Year 1990)
Budget Amount *help
¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 1990: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1989: ¥1,300,000 (Direct Cost: ¥1,300,000)
KeywordsPlasmid / Clustered inversion region / Shufflon / DNA rearrangement / Conjugal transfer
Research Abstract

Shufflon, or clustered inversion region, is a novel type of DNA rearrangement found in the IncI1 plasmid R64. The shufflon of R64 consists of four DNA segments, designated A, B, C and D, which differ in DNA length and sequence. The four DNA segments are flanked and separated by seven 19-bp repeat sequences. Site-specific recombination between any two inverted repeats results in a complex DNA rearrangement. An analysis of open reading frames revealed that the shufflon is a biological switch to select one of the seven C-termini of a gene, pilV.
By constructing a series of deletion-derivatives of plasmid R64, a region responsible for the conjugal transfer was located within a 54-kb DNA region to the right of replication region. We determined the nucleotide sequence of a 19-kb DNA region responsible for the formation of thin pilus, and found that 18 open reading frames (ORFs) are present in the same orientation. The pliv gene is the last one of the 18 ORFs. By introducing deletion and inser … More tion mutations into all the 18 ORFs, we demonstrated that most of the 18 ORFs are required for the formation of thin pilus and for the conjugal transfer. To reveal the role of the DNA rearrangement in the shufflon region, we constructed R64 derivatives where the C-termini of the pilv gene are fixed. Donor cells carrying R64 derivative plasmids with the seven fixed pilv C-termini were mated with one of several recipient bacterial strains including E. Coli K-12, B and C, Salmonella typhimurium LT2 and Shigella flexneri. The results indicated that the transfer frequencies of liquid mating dramatically change according to the combination of recipient strains and variable C-terminiof the pilv gene in the donors, suggesting that the shufflon determines the recipient specificity in liquid mating in plasmid R64. We also sequenced the oriT region located at the other end of transfer region of R64 and found two open reading frames, designated nikA and nikB. NikA and nikB were found indispensable for the transfer of R64 and were demonstrated to function together as an oriT-specific nickase. Less

Report

(3 results)
  • 1990 Annual Research Report   Final Research Report Summary
  • 1989 Annual Research Report
  • Research Products

    (25 results)

All Other

All Publications (25 results)

  • [Publications] Teruya Komano: "Shufflon:multiーinversion of four contiguous DNA segments of plasmid R64 creates seven different open reading frames." Nucleic Acids Res.15. 1165-1172 (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Akihiro Kubo: "Nucleotide sequence of the rci gene encoding shufflonーspecific DNA recombinase in the IncI1 plasmid R64:Homology to the siteーspecific recombinases of integrase family." Mol.Gen.Genet.213. 30-35 (1988)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Morimasa Wada: "Participation of the hup gene products in siteーspecific DNA inversion in Escherichia coli." Gene. 76. 345-352 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] SuーRyang Kim: "Cloning and nucleotide sequence of the colIb shufflon." Plasmid. 22. 180-184 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Teruya Komano: "Transfer region of IncI1 plasmid R64 and role of shufflon in R64 transfer." J.Bacteriol.172. 2230-2235 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Teruya Komano: "Physical and genetic analyses of IncI2 plasmid R721:Evidence for the presence of shufflon." Plasmid. 23. 248-251 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Komano, T., A. Kubo, & T. Nisioka. Shufflon: "multi-inversion of four contiguous DNA segments of plasmid R64 creates seven different open reading frames." Nucl. Acids Res. 15. 1165-1172 (1987)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Kubo, A., A. Kusukawa, & T. Komano: "Nucleotide sequence of the rci gene encoding shufflon-specific DNA recombination in the IncIl plasmid R64." Mol. Gen. Genet.213. 30-35 (1988)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Komano, T., A. Toyoshima, K. Morita, & T. Nisioka: "Cloning and nucleotide sequence of the oriT region of the IncI1 plasmid R64." J. Bacteriol.170. 4385-4387 (1988)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Wada, M., K. Kutsukake, T. Komano, F. Imamoto, & Y. Kano: "Participation of the hup gene product in site-specific DNA inversion in Escherichia coli." Gene. 76. 345-352 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Kim, S. R., & T. Komano: "Cloning and nucleotide sequence of the ColIb shufflon." Plasmid. 22. 180-184 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Komano, T., N. Funayama, S. R. Kim, & T. Nisioka: "Transfer region of IncI1 plasmid R64 and role of shufflon in R64 transfer." J. Bacteriol.172. 2230-2235 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Komano, T., S. Fujitani, N. Funayama, A. Kanno, & K. Sakuma: "Physical and genetic analyses of IncI2 plasmid R721 : Evidence for the presence of shufflon." Plasmid. 23. 248-251 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Teruya Komano: "Shufflon:multiーinversion of four contiguous DNA segments of plasmid R64 creates seven different open reading frames." Nucleic Acids Res.15. 1165-1172 (1987)

    • Related Report
      1990 Annual Research Report
  • [Publications] Akihiro Kubo: "Nucleotide sequence of the rci gene encoding shufflonーspecific DNA recombinase in the IncI1 plasmid R64:Homology to the siteーspecific recombinases of integrase family." Mo1.Gen.Genet.213. 30-35 (1988)

    • Related Report
      1990 Annual Research Report
  • [Publications] Morimasa Wada: "Participation of the hup products in siteーspecific DNA inversion in Escherichia coli." Gene. 76. 345-352 (1989)

    • Related Report
      1990 Annual Research Report
  • [Publications] SuーRyang Kim: "Cloning and nucleotide sequence of the ColIb shufflon." Plasmid. 22. 180-184 (1989)

    • Related Report
      1990 Annual Research Report
  • [Publications] Teruya Komano: "Transfer region of IncI1 plasmidR64 and role of shufflon in R64 transfer." J.Bacteriol.172. 2230-2235 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Teruya Komano: "Physical and genetic analyses of IncI2 plasmid R721: Evidence for the presence of shufflon." Plasmid. 23. 248-251 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Teruya Komano: "Shufflon:multi-inversion of four contigous DNA segments of plasmid R64 creates seven different open reading frames." Nucleic Acids Res.15. 1165-1172 (1987)

    • Related Report
      1989 Annual Research Report
  • [Publications] Akihiro Kubo: "Nucleotide sequence of the rci gene encoding shufflon-specific DNA recombinase in the IncI1 plasmid R64:Homology to the site-specific recombinases of integrase family." Mol.Gen.Genet.213. 30-35 (1988)

    • Related Report
      1989 Annual Research Report
  • [Publications] Teruya Komano: "Cloning and nucleotide sequence of the oriT region of the IncI1 plasmid R64." J.Bacteriol.170. 4385-4387 (1988)

    • Related Report
      1989 Annual Research Report
  • [Publications] Morimasa Wada: "Participation of the hup gene products in site-specific DNA inversion in Escherichia coli." Gene. 76. 345-352 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] Su-Ryang Kim: "Cloning and nucleotide sequence of the ColIb shufflon." Plasmid. 22. 180-184 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] Teruya Komano: "Transfer region of IncI1 plasmid R64 and role of shufflon in R64 transfer." J.Bacteriol.172. (1990)

    • Related Report
      1989 Annual Research Report

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Published: 1989-04-01   Modified: 2016-04-21  

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