Mechanisms of transcription in eosinophils

GATA-1, but not GATA-2, transactivates the promoter of the eosinophil granule major basic protein gene

Yuji Yamaguchi, Steven J. Ackerman, Naoko Minegishi, Masaki Takiguchi, Masayuki Yamamoto, Toshio Suda

Research output: Contribution to journalArticle

56 Citations (Scopus)

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 3.2 kb of sequence upstream of the exon 9 transcriptional start site (P2 promoter) and serial 5' deletions into the pXP2 luciferase reporter vector. An 80% decrement in promoter activity was obtained when MBP sequences between bp -117 to -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 50-bp region between the two genes. To determine which GATA proteins bind this consensus site, we performed electrophoretic mobility shift assays (EMSAs), which showed 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 combination, can transactivate the MBP promoter in the Jurkat T cell line. Cotransfection with a GATA-1 expression vector produced 20-fold augmentation of MBP promoter activity, whereas GATA-2 had no activity. In contrast, combined cotransfection 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, a negative regulatory role for GATA-2 in MBP expression, and possibly eosinophil gene transcription in general during myelopoiesis.

Original languageEnglish
Pages (from-to)3447-3458
Number of pages12
JournalBlood
Volume91
Issue number9
Publication statusPublished - 1998 May 1

Fingerprint

Eosinophil Major Basic Protein
Transcription
Eosinophils
Genes
Proteins
Transcription Factors
Myelopoiesis
Electrophoretic mobility
Jurkat Cells
T-cells
Basophils
Trophoblasts
Electrophoretic Mobility Shift Assay
Binding sites
Luciferases
Exons
Assays
Binding Sites

ASJC Scopus subject areas

  • Hematology

Cite this

Yamaguchi, Y., Ackerman, S. J., Minegishi, N., Takiguchi, M., Yamamoto, M., & Suda, T. (1998). Mechanisms of transcription in eosinophils: GATA-1, but not GATA-2, transactivates the promoter of the eosinophil granule major basic protein gene. Blood, 91(9), 3447-3458.

Mechanisms of transcription in eosinophils : GATA-1, but not GATA-2, transactivates the promoter of the eosinophil granule major basic protein gene. / Yamaguchi, Yuji; Ackerman, Steven J.; Minegishi, Naoko; Takiguchi, Masaki; Yamamoto, Masayuki; Suda, Toshio.

In: Blood, Vol. 91, No. 9, 01.05.1998, p. 3447-3458.

Research output: Contribution to journalArticle

Yamaguchi, Y, Ackerman, SJ, Minegishi, N, Takiguchi, M, Yamamoto, M & Suda, T 1998, 'Mechanisms of transcription in eosinophils: GATA-1, but not GATA-2, transactivates the promoter of the eosinophil granule major basic protein gene', Blood, vol. 91, no. 9, pp. 3447-3458.
Yamaguchi Y, Ackerman SJ, Minegishi N, Takiguchi M, 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. 1998 May 1;91(9):3447-3458.
Yamaguchi, Yuji ; Ackerman, Steven J. ; Minegishi, Naoko ; Takiguchi, Masaki ; Yamamoto, Masayuki ; Suda, Toshio. / Mechanisms of transcription in eosinophils : GATA-1, but not GATA-2, transactivates the promoter of the eosinophil granule major basic protein gene. In: Blood. 1998 ; Vol. 91, No. 9. pp. 3447-3458.
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