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1997 Fiscal Year Final Research Report Summary

Isolation of genes involved in the commitment of eosinophils and basophils

Research Project

Project/Area Number 08671244
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Hematology
Research InstitutionKumamoto University

Principal Investigator

YAMAGUCHI Yuji  Kumamoto University School of Medicir Lecturer, 医学部, 講師 (00220286)

Project Period (FY) 1996 – 1997
Keywordseosinophils / differentiation / Transcription factors / promoter / GATA-1 / GATA-2
Research Abstract

Granule major basic protein (MBP), is expressed exclusively in eosinophils, basophils, and placental trophoblasts. To identify the cis-elements and transcription factors involved in regulating MBP expression, we subcloned 3.2kb of MBP upstream sequence and serial 5' deletions into the pXP2 luciferase reporter vector. An 80% decrement in promoter activity was obtained when MBP sequences between bp -117 and -67 were deleted. To identify transcription factors that bind to and transactivate through the bp -117 to -67 region, we first compared the upstream genomic sequences of human and murine MBP ; a potential GATA binding consensus site was conserved in the 50bp region between the two genes. To determine which GATA proteins bind this consensus site, we performed electrophoretic mobility shift assays (EMSAs) which demonstrated that both GATA-1 and GATA-2 can bind to this consensus site. To determine the functionality of this site, we tested whether GATA-1 and GATA-2, either individually or in combinantion, can transactivate the MBP promoter in the Jurkat T cell line. Co-transfection with a GATA-1 expression vector produced 20-fold augmentation of MBP promoter activity, whereas GATA-2 had no activity. In contrast, combined co-transfection of GATA-1 and GATA-2, decreased the ability of GATA-1 to transactivate the MBP promoter by approximately 50%. Our results provide the first evidence for a GATA-1 target gene in eosinophils, and a negative regulatory role for GATA-2 in MBP expression, and possibly eosinophil gene transcription in general during myelopoiesis.

  • Research Products

    (8 results)

All Other

All Publications (8 results)

  • [Publications] Yamaguchi Y: "Functional characterization of the promoter for the gene encoding murhe CD34" Biochim.Biophys.Acta. 1350. 141-146 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Sonoda Y: "Stimulation of interleukin-8 production by Okadaic acid and vanadate in a human pronyelo cytic cell line,an HL-60 subline" J.Biol.Chem.272. 15366-15372 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yamaguchi Y: "Forced GATA-1 expression in the marine cell line Ml:Induction of c-mpl exression and megakaryo cytic erythroid differentation" Blood. 91. 450-457 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yamaguchi Y: "Mechanisms of trans cription in eosinophils;GATA-1,but not GATA-2 fransactivates the promoter of the eosinephil granule major basic protreingene" Blood. (in press). (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yamaguchi, Y., Tenen, D.G., and Suda, T.: "Functional Characterization of the Promoter for the Gene Encoding Murine CD34." Biochim.Biophys.Acta. 1350. 141-146 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Sonoda, Y., Kasahara, T., Yamaguchi, Y., Kuno, K., Matsushima, K., Mukaida, N.: "Stimulation of interleukin-8 production by Okadaic acid and vanadate in a human promyelocytic cell line, an HL-60 subline." J.Biol.Chem.272. 15366-15372 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yamaguchi, Y., Zon, L.I., Ackerman, S.J., Yamamoto, M., Suda, T.: "Forced GATA-1 expression in the murine cell line M1 : Induction of c-mpl expression and megakaryocytic/erythroid differentiation." Blood. 91. 450-457 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yamaguchi, Y., Ackerman, S.J., Minegishi, N., Yamamoto, M., Suda, T.: "Mechanisms of transcription in eosinophils : GATA-1, but not GATA-2, transactivates the promoter of the eosinophil granule major basic protein gene." Blood. (in press). (1998)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 1999-03-16  

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