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Interaction of protcin and DNA by AFM (Analysis of the bacterial stress responsive factors binding to DNA by Atomic Force Microscopy).

Research Project

Project/Area Number 09044254
Research Category

Grant-in-Aid for international Scientific Research

Allocation TypeSingle-year Grants
SectionJoint Research
Research Field Bacteriology (including Mycology)
Research InstitutionUniversity of Tsukuba

Principal Investigator

OHTA Toshiko  College of Medical Technology and Nursing Professor, 医療技術短期大学部, 教授 (40233134)

Co-Investigator(Kenkyū-buntansha) PATEL Smia.S.  Ohio State University Biochemistry Asociate Professor, 生物科学科生化学研究室, 助教授
HAI Tsonwin  Ohio State University Neuroscience Center Asociate Professor, 神経生物工学センター, 助教授
KURAZONO Hisao  University of Tsukuba Microbiology Asistant Professor, 基礎医学系, 講師 (90186487)
HAYASHI Hideo  University of Tsukuba Microbiology Professor, 基礎医学系, 教授 (40033203)
TAKEYASU Kunio  Kyoto University Integrated Human Studies Professor, 人間科学部, 教授 (40135695)
SMITA S Pate  オハイオ州立大学, 生物科学科・生化学研究室, 助教授
TSONWIN Hai  オハイオ州立大学, 神経生物工学センター, 助教授
Project Period (FY) 1997
Project Status Completed (Fiscal Year 1997)
Budget Amount *help
¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 1997: ¥3,400,000 (Direct Cost: ¥3,400,000)
KeywordsAtomic Force Microscopy / Staphylococcus aureus / Heat shock protein / Stress response / Regulator / CIRCE element
Research Abstract

(1) Disruption and complcmentation of hrc37 of the HSP70 operon in Staphylococcus aureus showed that the transcripts from those CIRCE (Controlling lnvertcd Repeat of Chaperon Expression) depcndcnt promoters were repressed by Hrc37 over-products ; however transcripts from the CIRCE independent promoters were not affected in this case. (submitted)
(2) Furthermore, gel retardation assay and Atomic Force Microscopy (AFM) with a specific CIRCE probe revealed that the Hrc37 could be involved in CIRCE dependent repression of heat induction.
(3) Sigma factor sigB-knockout mutant revealed that SigB enhanced expression of alkaline responsible asp genes, pathogenic factor-controlled gene, sar, and carotenoid-producing gcncs, crtM/N at stationary phase. The SigB recognized a specific sequence, GGGTAT in the promoters of these genes.
(4) Using differential hybridization analysis, one of the S.aureus genomic library were positively induced by heat, cadmium, mercury and zinc insults. DNA sequencing and … More Northern analysis revealed that 1.4kb transcript containing two open reading frames (czrA and czrB) as induced by heat and metal insults. The CzrA may play a repressor role in order to autoregulate its own operon. (submitted)
(5) We found a novel 35-kDa membrane protein induced especially by antibiotics that affect S.aureus cell wall. Analysis of SDS-PAGE revealed that a 35-kDa protein band was evidently enhanced after 30min in the presence of beta-lactams. The gene was cloned by differential hybridization with mRNAs enhanced in the prcsence of beta-lactams. The enzymatic activities of the gene were analyzed to make clear its functional role.
(6) A novel transcriptional factor ATF4 was purified and characterized biochemically. When ATF4 was reacted with CREB protein, the complex bound an element, which controlled Gadd153. (published)
(7) Structural analyzes of membrane proteins and nucleic acids by atomic force microscopy (AFM) have demonstrated that channel-like structures exist in both ion channels and ion pumps ; and that distinct higher-order structures such as stem-loop can be formed in the promoter regions of genes. (submitted) Less

Report

(2 results)
  • 1997 Annual Research Report   Final Research Report Summary
  • Research Products

    (20 results)

All Other

All Publications (20 results)

  • [Publications] 太田 敏子: "グラム陽性菌のストレス応答機構と調節系" 生化学. 71(1). 36-42 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Toshiko Ohta: "Structural and Functional Characterization of Homo-oligomeric Complexes of a and β Chaperonin Subunits fromthe Hyperthermophilic Archaeum Thermococcus strain KS-1" J.Mol.Biol.273. 635-645 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Toshiko Ohta: "HSP70 and HSP60 Operons are Regulated by a Transcriptional factor Hrc37 of the HSP70 Operon in Staphylococcus aureus" J.Bacteriol.(in Press). (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Toshiko Ohta: "Molecular Assembly of Biological Macromolecules Studied by Atomic Force Microscopy:Quantitative analyses ofmembrane structures and DNA-protein interactions" Annalytical Bioscience.(in Press). (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Tsonwin Hai: "The hepatitis B virus X proteinenhances the DNA-binding potential andtranscription efficacy of bZip transcription factors" J.Biol.Chem. 272. 20684-20690 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Tsonwin Hai: "Characterization of ATF4:a transcriptional activator that interacts with multiple domains of CBP" J.Biol.Chem. 272. 24088-24095 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Yoshida T., Yohda M., Iida T., Maruyama T., Taguchi H., Yoshida M., Yazaki K., Ohta T., Endo I., and Kagawa Y.: "Molecular cloning and expression of group II chaperonin from the hyperthermophilicarchaeum, Thermococcus strain KS-1 : the homooligomeric complexes ofrecombinant alpha and betasubunits have chaperonin activity." J.Biol.Chem.271. 22017-22021 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Takeyasu K., Tokumasu F., Yoshimura S., Ohta T., Omote H., Futai M., Maruyama K., and Sato M.: "Molecular assembly of biological macromolecules studied by Atomic Force Microscopy : Quantitative analyzes of membrane structures and DNA-protein interactions." Analytical Bioscience. (in press). (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Yoshida T., Yohda M., Iida T., Maruyama T., Taguchi H., Yazaki K., Ohta T., Odaka M., Endo I., and Kagawa Y: "Structural and functional characterization of homo-oligomeric complexes of a and b chaperonin subunits from the hyperthermophilic Archaeum Thermococcus strain KS-1." J.Mol.Biol.273 (3). 635-645 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kuroda M., Kobayashi D., Honda K., Hayashi H., and Ohta T.: "HSP70 and HSP60 operons are regulated by a transcriptional factor Hrc37 of the HSP70 operon in Staphylococcus aureus." J.Bacteriol.(in press). (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Hai S.et al.: "The hepatitis B virus X protein enhances the DNA-binding potential and transcription efficiency of bZip transcriptionfactors." J.Biol.Chem.272. 20684-20690 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] HaiS.et al: "Characterization of ATF4 : a transcriptional activator that interacts with multiple domains of CBP." J.Biol.Chem.272. 24088-24095 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 太田 敏子: "グラム陽性菌のストレス応答機構と調節系" 生化学. 71 (1). 36-42 (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] Toshiko Ohta: "Structural and Functional Characterization of Homo-oligomeric Complexes of α and β Chaperonin Subunits from the Hyperthermophilic Achaeum Thermococcus strain KS-1" J. Mol. Biol.273. 635-645 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Toshiko Ohta: "HSP70 and Operons are Regulated by a Transcriptional factor Hrc37 of the HSP70 Operon in Staphylococcus aureus" J. Bacteriol.(in Press). (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] Toshiko Ohta: "Molecular Assembly of Biological Macromolecules Studied by Atomic Force Microscopy : Quantitative analyses of membrane structures and DNA-protein interactions" Annalytical Bioscience.(in Press). (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] Tsonwin Hai: "The hepatitis B virus X protein enhances the DNA-binding potential and transcription efficacy of bZip transcription factors" J. Biol. Chem.272. 20684-20690 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Tsonwin Hai: "Characterization of ATF4 : a transcriptional activator that interacts with multiple domains of CBP" J. Biol. Chem.272. 24088-24095 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Tsonwin Hai: "Gadd153/Chop10, a potential target gene of the transcriptional repressor ATF3" Mol. Cell. Biol.17. 6700-6707 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Tsonwin Hai: "EGF induction of the c-jun promoter by a Rac pathway" Mol. Cell. Biol.18. 1065-1073 (1998)

    • Related Report
      1997 Annual Research Report

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Published: 1997-04-01   Modified: 2016-04-21  

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