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Analyses of cytokine gene expression by helper T cell subsets : role of NFAT-mediated gene activation and subset-specific regulatory mechanism.

Research Project

Project/Area Number 08457103
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Immunology
Research InstitutionThe University of Tokyo

Principal Investigator

ARAI Ken-ichi  Inst.of Medical Science, The University of Tokyo, Professor, 医科学研究所, 教授 (00012782)

Co-Investigator(Kenkyū-buntansha) SATO Noriko  Inst.of Medical Science, The University of Tokyo, Assistant Professor, 医科学研究所, 助手 (70280956)
WATANABE Sumiko  Inst.of Medical Science, The University of Tokyo, Assistant Professor, 医科学研究所, 助手 (60240735)
MASAI Hisao  Inst.of Medical Science, The University of Tokyo, Associate Professor, 医科学研究所, 助教授 (40229349)
YOKOTA Takashi  Inst.of Medical Science, The University of Tokyo, Professor, 医科学研究所, 客員教授 (50134622)
Project Period (FY) 1996 – 1998
Project Status Completed (Fiscal Year 1998)
Budget Amount *help
¥8,100,000 (Direct Cost: ¥8,100,000)
Fiscal Year 1998: ¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 1997: ¥2,600,000 (Direct Cost: ¥2,600,000)
Fiscal Year 1996: ¥2,500,000 (Direct Cost: ¥2,500,000)
KeywordsT lymphocytes / Cytokines / Helper T cell subsets / Transcription factors / Nuclear translocation / NFAT / GATA-3 / Chromatin structure / NF-AT / DNaseI hypersensitive site / NF-KB / NF-κB
Research Abstract

The aim of this study is to elucidate the regulatory mechanism of cytokine gene expression in helper T cells and their subsets.
(1) NFAT : regulation and function in T-cell activation. We have isolated human and murine genes of NFAT, a transcription factor that plays a critical role in transcriptional induction of cytokine genes. Throughout the mapping study on NFATx, a member of the NFAT family, we have defined a series of functional domains ; Rd domain critical for specific DNA-binding and association with AP-1 proteins, N-terminal domain for calcineurin-regulated activation of NFAT via nuclear translocation, and C-terminal domain for transactivation. in N-terminal domain, one of the critical segment for control of subcellular localization was narrowed down to 60-residue motif in N-terminal domain, as designated CRI.By modification of NFATx molecule (e.g., by deleting CRI or modifying calcineurin-interacting domains), we generated constitutively active of dominant negative mutants of … More NFATx with calcineurin-independent subcellular localization, that should be a useful tools for further understanding of NFAT-mediated gene regulation. We further analyzed expression and function of the NFAT family in thymocytes in which NFATx is predominantly expressed. We found that NFAT family mRNAs are developmentally regulated during the differentiation of T cells in the thymus, and that NFATx protein selectively contributes to the calcineurin-dependent NFAT-DNA binding activity in the DP stage, suggesting that NFAT is also involved in the signals required for generation of T cells in the thymus.
(2) Mechanism of Th2-specific cytokine gene regulation. We have previously described the regulatory motives in the promoters of cytokine genes including GM-CSF, IL-2, IL-3, IL-4, and IL-5, leading to discovery of conserved lymphokine elements (CLE). In current study was analyzed factors that facilitate the Th-subset specific expression of cytokines. In the analyzes of IL-5 promoter, 1.2 Kb-long promoter region was shown to define transactivation of IL-5 gene only in Th2 clones. Extensive studies of this region lead to discovery of NFIL-5CLEO and NFIL-5C, and GATA-3 or related transcription factor was indicated to involved in Th2-specific IL-5 gene induction via IL-5C-driven transactivation. Furthermore, in case of IL-4, the proximal promoter gives considerably less activity than intact IL-4 gene locus in Th2 cells. We performed DNaseI hypersensitive site (HSS) analysis using Th clones and in vitro differentiated Th1/Th2 cells, and identified that two out of three HSS located in IL-4 - IL-13 intergenic region were Th2-specific and appeared only after differentiation. These results provide novel and multiple regulatory mechanism in differential expression of Th2 cytokine gees. Less

Report

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

    (29 results)

All Other

All Publications (29 results)

  • [Publications] Arai K.,et al.: "Cytokine signal networks and a new era in biomedical research." Mol.Cells. 7. 1-12 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Masuda E.S.,et al.: "Control of NFATxl nuclear translocation by a calcineurin-regulated inhibitory domain." Mol.Cell Biol.17. 2066-75 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Liu J.,et al.: "Calcineurin-dependent nuclear translocation of a murine transcription factor NFATx : molecular cloning and functional characterization." Mol.Biol.Cell. 8. 157-70 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Amasaki Y.,et al.: "Distinct NFAT family proteins are involved in the nuclear NFAT-DNA binding complexes from human thymocyte subsets." J.Immunol.160. 2324-33 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Lee HJ.,et al.: "Characterization of cis-regulatory elements and nuclear factors conferring Th2-specific expression of the IL-5 gene : a role for a GATA-binding protein." J.Immunol.160. 2343-52 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Takemoto N.,et al.: "Th2-specific DNase I-hypersensitive sites in the murine IL-13 and IL-4 intergenic region." Int.Immunol.10. 1981-5 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Arai K,Tsuruta L,Watanabe S,Arai N.: "Cytokine signal networks and a new era in biomedical research." Mol.Cells. 7. 7-12 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Masuda ES,Liu J,Imamura R,Imai S,Arai K,Arai N.: "Control of NFAx1 nuclear translocation by a calcineurin-regulated inhibitory domain." Mol.Cell Biol. 17. 2066-75 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Liu J,Koyano-Nakagawa N,Amasaki Y,Saito-Ohara F,Ikeuchi T,Imai S,Takano T,Arai N,Yokota T,Arai K.: "Calcineurin-dependent nuclear translocation of a murine transcription factor NFATx : molecular cloning and functional characterization." Mol.Biol Cell. 8. 157-70 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Amasaki Y,Masuda ES,Imamura R,Arai K,Arai N: "Distinct NFAT family proteins are involved in the nuclear NFAT-DNA binding complexes from human thymocyte subsets." J Immunol. 160. 2324-33 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Lee HJ,O'Garra A,Arai K,Arai N: "Characterization of cis-regulatory elements and nuclear factors conferring Th2-specific expression of the IL-5 gene : a role for a GATA- binding protein." J Immunol. 160. 2343-52 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Takemoto N,Koyano NN,Arai N,Arai K,Yokota T: "Four P-like elements are required for optima transcription of the mouse IL-4 gene : involvement of a distinct set of nuclear factor of activated T cells and activator protein-1 family proteins." Int Immunol. 9. 1329-38 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Takemoto N, et al.: "Th2-specific DNase I-hypersensitive sites in the murine IL-13 and IL-4 intergenic region." Int Immunol. 10. 1981-1985 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Imamura R, et al.: "Carboxyl-terminal 15-amino acid sequence of NFATxl is possibly created by tissue-specific splicing and is essential for transactivation activity in T cells" J.Immunol. 161. 3455-3463 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Amasaki Y, et al.: "Distinct NFAT family proteins are involved in the nuclear NFAT-DNA binding complexes from human thymocyte subsets." J.Immunol. 160. 2343-2352 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Masuda ES, et al.: "Signalling into the T-cell nucleus : NFAT regulation" Cell Signal. 10. 599-611 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Masuda ES, et al.: "Control of NFATxl nuclear translocation by a calcineurin-regulated inhibitory domain" Mol Cell Biol. 17. 2066-2075 (1997)

    • Related Report
      1998 Annual Research Report
  • [Publications] Liu J, et al.: "Calcineurin-dependent nuclear translocation of a murine transcription factor NFATx : molecular cloning and functional characterization" Mol Biol Cell. 8. 157-170 (1997)

    • Related Report
      1998 Annual Research Report
  • [Publications] Lee,H.J.et al: "Characterization of cis-regulatory elements and nuclear factors conferring Th-2 specific expression of the IL-5 gene : a role for a GATA-binding protein." J.Immunology. (in press).

    • Related Report
      1997 Annual Research Report
  • [Publications] Amasaki,Y.et al: "Distinict NFAT-family proteins are involved in the stimulation-dependent NFAT complexes from thymocyte subsets." J.Immunology. (in press).

    • Related Report
      1997 Annual Research Report
  • [Publications] Masuda,E.S.et al: "Control of NFATxl nuclear translocation by a calcineurin-regulated inhibitory domain." Mol.Cell.Biol.17.4. 2066-2075 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Takemoto,N.et al: "Four P like elements are required for optimal transcription of the mouse IL-4 gene : involvement of distinict set of NF-AT and AP-1 family proteins." Int.Immunol.9・9. 1329-1338 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Pan,S.et al: "Molecular cloning and functional characterization of murine cDNA encoding transcription factor NFATc." Biochem.Biophys.Res.Communi.240・2. 314-323 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Arai,K.et al: "Cytokine signal networks and a new era in biomedical research." Mol.Cells.7. 1-12 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Koyano-Nakagawa,N.,et al.: "Specific versus cooperative regulatory mechanism of the cytokine genes that are clustered on the same chromosome." J.Allergy Clinic.Immunol.98. 174-182 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Naito,Y.,et al: "Signal transduction in Th clones:Target of differential modulation by PGE2 may reside downstream of the PKC dependent pathway." Cytokine Research. 8. 346-356 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Harada,Y.,et al.: "Factors affecting the cytokine production of human T cells stimulated via different modes of activation." J.Allergy Clinic.Immunol.98. 161-173 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Liu,J.,et al.: "Calcineurin dependent nuclear translocation of a murine transcription factor NFATc:molecular clonning and functional characterization." Mol.Biol.Cell. 8. 157-170 (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] Masuda,E.S.,et al.: "NFATx1 Regulation by a calcium-dependent gate." Mol.Cell.Biol.,in press.(1997)

    • Related Report
      1996 Annual Research Report

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

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