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CONTROL OF INITIATION FREQUENCY OF COLE2 REPLICATION

Research Project

Project/Area Number 08458217
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Molecular biology
Research InstitutionSHINSHU UNIVERSITY

Principal Investigator

ITOH Tateo  SHINSHU UNIVERSITY,FACULTY OF SCIENCE,PROFESSOR, 理学部, 教授 (40051817)

Co-Investigator(Kenkyū-buntansha) KUBO Hiroyoshi  SHINSHU UNIVERSITY,FACULTY OF SCIENCE,LECTURER, 理学部, 講師 (60205127)
Project Period (FY) 1996 – 1998
Project Status Completed (Fiscal Year 1998)
Budget Amount *help
¥7,700,000 (Direct Cost: ¥7,700,000)
Fiscal Year 1998: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1997: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1996: ¥5,100,000 (Direct Cost: ¥5,100,000)
KeywordsColE2 plasmid / Replication initiator / primer RNA / origin / DNA binding protein / antisense RNA / translation / RNA secondary structure / ColE2プラスミド / 複製開始蛋白質 / 過剰複製阻止 / コピー数調節 / ColE2群プラスミド / 二量体形成 / 直列繰り返し配列 / RNaseIII / 複製開始頻度
Research Abstract

The ColE2 Rep protein is unique among many essential initiator proteins in that it is a plasmid-specified primase specific for initiation of DNA replication in the origin by DNA polymerase I.Expression of the Rep protein is kept constant through negative regulation by a small antisense RNA (RNAI). RNAl is entirely complementary to the 5' nontranslated region of the Rep mRNA.Since the Rep protein is trans-acting, it is not so obvious how the initiation frequency can be kept constant.
Analyses of various derivatives of the ColE2 origin carrying deletions or single-base substitutions showed that the region may be divided into three subregions : one important for stable binding of the Rep protein, another important for binding and for replication and another important for replication but apparently not for binding. Transformation frequency of the autonomously replicating plasmids carrying deletions in the first region is lower and nevertheless the copy numbers of them in hostbacteria are higher as compared with the wild-type plasmid. The Rep protein might inhibit over-replication or re-replication of newly replicated daughter molecules by stable binding to the origin. This might be important to keep the initiation frequency at a constant level.
The 5' nontranslated region of the Rep mRNA far upstream of the initiation codon and another region in the coding region near the initiation codon are important for efficient expression of the Rep protein. Partial digestion with various RNases in the presence or absence of RNAI suggested that binding of RNAI to the Rep mRNA causes changes in the secondary structure of the region containing the initiation codon. Mutant plasmids carrying base substitutions in the upstream and downstream regions of the initiation codon of the Rep mRNA have been isolated. Some of the mutations abolished or decreased expression of the Rep protein.

Report

(4 results)
  • 1998 Annual Research Report   Final Research Report Summary
  • 1997 Annual Research Report
  • 1996 Annual Research Report
  • Research Products

    (13 results)

All Other

All Publications (13 results)

  • [Publications] Miki Shinohara: "Specificity determinants in interaction of the initiator (Rep) proteins with the origins in the plasmids Co1E2-P9 and Co1E3-CA38 identified by chimera analysis." J.M.B.257. 290-300 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Masaru Yagura: "Functional prganization of plasmid Co1E2 initiator (Rep) protein and replication origin" Genes Genet.Syst.72. 382 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] 伊藤建夫(共著): "21世紀への遺伝学II 分子遺伝学(三浦謹一郎編) 3.遺伝子複製のメカニズム" 吉野達治(裳華房), 276 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] SHINOHARA,MIKI: "Specificity determinants in interaction of the initiator (Rep) proteins with the origins in the plasmids ColE2-P9 and ColE3-CA38 identified by chimera analysis." JMB. 257. 290-300 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] YAGURA,MASARU: "Functional organization of plasmid ColE2 initiator (Rep) protein and replication origin." GENES GENET.SYST.72. 382 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] ITOH,TATEO,YOSHINO,TATUJI: "Mechanism of DNA replication : in Genetics in the 21st century, II,molecular Genetics" 1997. 276

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Miki Shinohara: "Specificity determinants in interaction of the initiator (Rep) proteins with the origins in the plasmids ColE2-P9 and ColE3-CA38 identified by chimera analysis." J.M.B.257. 290-300 (1996)

    • Related Report
      1998 Annual Research Report
  • [Publications] Masaru Yagura: "Functional prganization of plasmid ColE2 initiator (Rep) protein and replication origin" Genes Genet Syst.72. 382 (1997)

    • Related Report
      1998 Annual Research Report
  • [Publications] 伊藤建夫 (共著): "21世紀への遺伝学II 分子遺伝学 (三浦謹一郎編)△3.遺伝子複製のメカニズム" 吉野達冶 (裳華房), 276 (1997)

    • Related Report
      1998 Annual Research Report
  • [Publications] Miki Shinohara: "Specificity determinants in interaction of the initiator(Rep)proteins with the origins in the plasmids ColE2-P9 and ColE3-CA38 identified by chimera analysis." J.M.B.257. 290-300 (1996)

    • Related Report
      1997 Annual Research Report
  • [Publications] Masaru Yagura: "Functional prganization of plasmid ColE2 initiator(Rep)protein and replication origin" Genes Genet.Syst.72. 382 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 伊藤建夫(共著): "21世紀への遺伝学II 分子遺伝学(三浦謹一郎編)3.遺伝子複製のメカニズム" 吉野達治(裳華房), 276 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Miki Shinohara: "Specificity deteminants in interaction of the initiator (Rep) proteins with the origins in the plasmids ColE2-P9 and ColE3-CA38 identified by chimera analysis." J.M.B.257. 290-300

    • Related Report
      1996 Annual Research Report

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Published: 1996-04-01   Modified: 2016-04-21  

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