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Studies on the regulation of DNA replication of Rtsl and P

Research Project

Project/Area Number 06044088
Research Category

Grant-in-Aid for international Scientific Research

Allocation TypeSingle-year Grants
SectionJoint Research
Research InstitutionShinshu University

Principal Investigator

TERAWAKI Yoshiro  Shinshu University, School of Medicine professor, 医学部, 教授 (10014333)

Co-Investigator(Kenkyū-buntansha) KAJI Akira  University of Pennsylvania, USA Research Assoicate, 医学部(米国), 教授
AUSTIN S.  国立癌研究所(米国), 部長
TABUCHI Akira  Shinshu University, School of Agriculture professor, 農学部, 助手 (50236725)
OHNISHI Makoto  Shinshu University, School of Medicine Director, 医学部, 助手 (10233214)
AUSTIN Stuart  National Cancer Institute, USA Research Assoicate
ABELES A.  国立癌研究所, 染色体研究部(米国), 主任研究員
Project Period (FY) 1994 – 1995
Project Status Completed (Fiscal Year 1995)
Budget Amount *help
¥4,000,000 (Direct Cost: ¥4,000,000)
Fiscal Year 1995: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 1994: ¥2,600,000 (Direct Cost: ¥2,600,000)
Keywordsplasmid Rts1 / initiator protein / hybrid protein / phage P1 / functional domain / DNA binding protein / hig phenotype / higA and higB / killer and suppressor / Phage P1 / Initiator protein / Hybrid protein / Initiation / Autorepression / DNA binding / Replication inhibition
Research Abstract

A.The joint research with Dr.S.Austin group
The Rtsl RepA protein, consisting of 288 amino acids, is a trans-acting protein essential for initiation of plasmid replication. To study the functional domains of RepA, hybrid proteins of Rtsl RepA with the initiator protein of phage P1 were constructed such that the N-terminal portion was from Rts1 RepA and the C-terminal portion was from P1 RepA (RepAXn), and the N-terminal from P1 RepA and the C-terminal from Rts1 RepA (RepALXn). We already reported the RepAXn functions in J.Bacteriol. 177 : 4028-4035, 1995, suggesting that the aminoacid residues 113 to 129 was important for ori binding in vitro, and the residues 177 to 206 region was required for ori activation in vivo as well as the ori binding domain.
Four RepALXn proteins were constructed ; RepALX12 (P1 RepA 256 aa/Rts1 RepA 31 aa), RepALX13 (205/82), RepALX15 (144/143), and RepALX17 (112/175) RepALX15 and RepALX17 activated Rts1 ori efficiently. This evidenced our previous finding that the 177 to 206 aa region is essential for Rts1 ori activation. The P1 ori was activated by RepALX12 and RepALX13, which suggests a quite similar domain structure of P1 RepA to rts1 RepA.
B.The joint research with Dr.A.Kaji group
We determined the locus related to the inhibition of the host cell growth on the Rts1 genome. The locus, named hig, consists of higA and higB genes, and their functions were analyzed. The HigB product (92 aa) displayd a killer activity to the host cells and the HigA protein (104 aa) suppressed the HigB function. The results will be appeared in Biochem. Biophys.Res.Commun. (in press).

Report

(2 results)
  • 1995 Final Research Report Summary
  • 1994 Annual Research Report
  • Research Products

    (6 results)

All Other

All Publications (6 results)

  • [Publications] 田渕晃: "Analysis of functional domains of Rts1 RepA by means of a series of hybrid proteins" Jounal of Bacteriology. 177. 4028-4035 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] 田慶宝: "A new plasmid encoded proteic killer gene system : cloning,sequencing and analysing hig--" Biochem.Biophys.Res.Commun.(in press). (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Akira Tabuchi: "Analysis of functional domains of Rts1 RepA by means of a series of hybrid proteins with P1 RepA" Journal of Bacteriology. 177. 4028-4035 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Quin Bao Tian: "A new plasmid encoded proteic killer gene system : coloning, sequencing and analysing hig locus of Rts1" Biochem.Biophys.Res.Commun.(in press). (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] 田渕,晃: "Rts1とP1とのhybrid蛋白の作成と機能的ドメインの解析" 日本細菌学雑誌. 49. 219 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Tabuchi,A.: "Analysis of functional domains of Rts1 RepA by constructing a series of hybrid proteins with P1 RepA" J.Bacteriol.(submitted).

    • Related Report
      1994 Annual Research Report

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Published: 1994-04-01   Modified: 2016-04-21  

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