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Comprehensive identification of nuclear receptor-regulating proteins usinghigh luminescent YFP-fusion cDNA library

Research Project

Project/Area Number 18591027
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Endocrinology
Research InstitutionKyushu University

Principal Investigator

KAWATE Hisaya  Kyushu University, Hospital, Assistant Professor (20336027)

Co-Investigator(Kenkyū-buntansha) TAKAYANAGI Ryoichi  Kyushu Unyversity, Graduate School of Medical Sciences, Professor (30154917)
Project Period (FY) 2006 – 2007
Project Status Completed (Fiscal Year 2007)
Budget Amount *help
¥3,890,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥390,000)
Fiscal Year 2007: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2006: ¥2,200,000 (Direct Cost: ¥2,200,000)
KeywordsNuclear receptor / Gene / Confocal microscope / Hormone
Research Abstract

1. Examination of the significance of nuclear foci formation of steroid hormone receptors We hound that ligand-dependent foci formation of steroid hormone receptors does not precede the transcriptional activation and depends on the relative expression ratio of coactivators and corepressors.
2.Domain analysis of transcription compressor N-CoR We generated various N-CoR deletion mutants and hound that the central portion (1133-1798 amino acid) of the protein was mostly responsible for both the interaction with steroid hormone receptors and the repression of their activities.
3.Mutual transactivational repression of Runx2 the androgen receptor (AR) Tb examine the role of steroid hormone receptors on bone metabolism, we analized mutual action between steroid hormone receptors and Runx2, which is a key regulator for osteoblast diffrentiation and proliferation. Our experimental results suggest that both AR and Runx2 repress the transcriptional function of the other protein by extracting it from its original compartment.
4.Construction of high luminescentYFP-cDNAlibrary and introdution into culture cells Messenger RNA was extracted from HeLa cells and cDNA was synthesized by reverse transcriptase and then fused to high luminescent YFP vector to make fusion library. This cDNA library was cotransfected with AR-expression plasmid into COS-7 cells. After the ligand treatment, we tried to collect cells showing specific nuclear patterns similar to those of AR. However, we failed to pick up such kind of cells because cells showing foci-like pattern was very rare.

Report

(3 results)
  • 2007 Annual Research Report   Final Research Report Summary
  • 2006 Annual Research Report
  • Research Products

    (21 results)

All 2008 2007 2006 Other

All Journal Article (6 results) (of which Peer Reviewed: 3 results) Presentation (13 results) Remarks (2 results)

  • [Journal Article] Methylation of a conserved intronic CpG island of mouse SF-1 is associatde with cell-specific expression of SF-1 in a culture system but not with tissue-specific expression2008

    • Author(s)
      Shirohzu H, et. al.
    • Journal Title

      Biochem Biophys Res Commun 369

      Pages: 862-867

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Mutual transactivational repression of Runx2 and the androgen receptor by an impairment of their normal compartmentalization2007

    • Author(s)
      Kawate H., et. al.
    • Journal Title

      J.Steroid Biochem.Mol.Biol 105

      Pages: 46-56

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Annual Research Report 2007 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] Mutual transactivational repression of Runx2 and the androgen receptor by an impairment of their normal compartmentalization.2007

    • Author(s)
      Kawate, H., et. al.
    • Journal Title

      J. Steroid Biochem. Mol. Biol. 105

      Pages: 46-56

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Nuclear compartmentalization of N-CoR and its interactions with steroid receptors2006

    • Author(s)
      Wu Y, et. al.
    • Journal Title

      Molecular and Cellular Biology 26

      Pages: 6633-6655

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] Nuclear compartmentalization of N-CoR and its interactions with steroid receptors2006

    • Author(s)
      Wu, Y, et. al.
    • Journal Title

      Molecular and Cellular Biology 26

      Pages: 6633-6655

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Nuclear compartmentalization of N-CoR and its interactions with steroid receptors2006

    • Author(s)
      Wu Y et al.
    • Journal Title

      Molecular and Cellular Biology 26・17

      Pages: 6633-6655

    • Related Report
      2006 Annual Research Report
  • [Presentation] Significance of ligand-induced intranuclear foci formation of steroid hormone receptors.2007

    • Author(s)
      Kawate, H
    • Organizer
      The Endocrine Society's 89th Annual Meeting
    • Place of Presentation
      Toronto, Canada
    • Year and Date
      2007-12-09
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Significance of ligand-induced intranuclear foci formation of steroid hormone receptors2007

    • Author(s)
      Hisaya Kawate
    • Organizer
      The Endocrine Society's 89^<th> Annual Meeting
    • Place of Presentation
      Toronto,Canada
    • Year and Date
      2007-06-04
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Significance of ligand-induced intranuclear foci formation of steroidhormone receptors2007

    • Author(s)
      Hisaya Kawate
    • Organizer
      The Endocrine Society's 89^<th> Annual Meeting
    • Place of Presentation
      Toronto, Canada
    • Year and Date
      2007-06-04
    • Related Report
      2007 Annual Research Report
  • [Presentation] N-CoR competitively suppresses the transactivation function of steroid hormone receptors with coactivators through its middle region2006

    • Author(s)
      Yin Wu
    • Organizer
      第14回日本ステロイドホルモン学会
    • Place of Presentation
      大阪
    • Year and Date
      2006-12-09
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] N-CoR competitively suppresses the transactivation function of steroid hormone receptors with coactivators through its middle region.2006

    • Author(s)
      Yin, Wu
    • Organizer
      14th Japanese Steroid Hormone Society
    • Place of Presentation
      Osaka
    • Year and Date
      2006-12-09
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Nucelar compartmentalization of N-CoR and steroid hormone receptors2006

    • Author(s)
      Yin Wu
    • Organizer
      日本分子生物学会2006フォーラム
    • Place of Presentation
      名古屋
    • Year and Date
      2006-12-08
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Nucelar compartmentalization of N-CoR and steroid hormone receptors.2006

    • Author(s)
      Yin, Wu
    • Organizer
      MBSJ 2006 Forum
    • Place of Presentation
      Nagoya
    • Year and Date
      2006-12-08
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Testicular Zinc Finger Protein Suppresses the Transactivation Function and Intranuclear Foci Formation of Agonist-Bound Androgen Receptor through Recruitment of Histone Deacetylase 22006

    • Author(s)
      Hisaya Kawate
    • Organizer
      The Endocrine Society's 88th Annual Meeting
    • Place of Presentation
      Boston,USA
    • Year and Date
      2006-06-25
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Ligand-Induced Intranuclear Compartmentalization of Androgen Receptor Is Dose Dependent and Coupled to Its Mediated Transactivation2006

    • Author(s)
      Yin Wu
    • Organizer
      The Endocrine Society's 88^<th> Annual Meeting
    • Place of Presentation
      Boston,USA
    • Year and Date
      2006-06-25
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Two Alternative Splicing Variants of Testicular Zinc Finger Protein Show Opposite Effects on Transactivation Function of Androgen Receptor2006

    • Author(s)
      Ryoichi Takayanagi
    • Organizer
      The Endocrine Society's 88^<th> Annual Meeting
    • Place of Presentation
      Boston,USA
    • Year and Date
      2006-06-25
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Testicular Zinc Finger Protein Suppresses the Trans-activation Function and Intranuclear Foci Formation of Agonist-Bound Androgen Receptor through Recruitment of Histone Deacetylase 2.2006

    • Author(s)
      Hisaya, Kawate
    • Organizer
      The Endocrine Society's 88th Annual Meeting
    • Place of Presentation
      Boston, USA
    • Year and Date
      2006-06-25
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Ligand-Induced Intranuclear Compartmentalization of Androgen Receptor Is Dose Dependent and Coupled to Its Mediated Transactivation.2006

    • Author(s)
      Yin, Wu
    • Organizer
      The Endocrine Society's 88th Annual Meeting
    • Place of Presentation
      Boston, USA
    • Year and Date
      2006-06-25
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Two Alternative Splicing Variants of Testicular Zinc Finger Protein Show Opposite Effects on Transactivation Function of Androgen Receptor2006

    • Author(s)
      Ryoichi, Takayanagi
    • Organizer
      The Endocrine Society's 88th Annual Meeting
    • Place of Presentation
      Boston, USA
    • Year and Date
      2006-06-25
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Remarks] 「研究成果報告書概要(和文)」より

    • URL

      http://hyoka.ofc.kyushu-u.ac.jp/search/index.html

    • Related Report
      2007 Final Research Report Summary
  • [Remarks]

    • URL

      http://hyoka.ofc.Kyushu-u.ac.jp/search/index.html

    • Related Report
      2007 Annual Research Report

URL: 

Published: 2006-04-01   Modified: 2016-04-21  

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