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The leukemia associated ETO nuclear repressor gene is regulated by the GATA-1 transcription factor in erythroid/megakaryocytic cells

Ajore, Ram LU ; Dhanda, Rakesh Singh ; Gullberg, Urban LU and Olsson, Inge LU (2010) In BMC Molecular Biology 11.
Abstract

BACKGROUND: The Eight-Twenty-One (ETO) nuclear co-repressor gene belongs to the ETO homologue family also containing Myeloid Translocation Gene on chromosome 16 (MTG16) and myeloid translocation Gene-Related protein 1 (MTGR1). By chromosomal translocations ETO and MTG16 become parts of fusion proteins characteristic of morphological variants of acute myeloid leukemia. Normal functions of ETO homologues have as yet not been examined. The goal of this work was to identify structural and functional promoter elements upstream of the coding sequence of the ETO gene in order to explore lineage-specific hematopoietic expression and get hints to function.

RESULTS: A putative proximal ETO promoter was identified within 411 bp upstream of... (More)

BACKGROUND: The Eight-Twenty-One (ETO) nuclear co-repressor gene belongs to the ETO homologue family also containing Myeloid Translocation Gene on chromosome 16 (MTG16) and myeloid translocation Gene-Related protein 1 (MTGR1). By chromosomal translocations ETO and MTG16 become parts of fusion proteins characteristic of morphological variants of acute myeloid leukemia. Normal functions of ETO homologues have as yet not been examined. The goal of this work was to identify structural and functional promoter elements upstream of the coding sequence of the ETO gene in order to explore lineage-specific hematopoietic expression and get hints to function.

RESULTS: A putative proximal ETO promoter was identified within 411 bp upstream of the transcription start site. Strong ETO promoter activity was specifically observed upon transfection of a promoter reporter construct into erythroid/megakaryocytic cells, which have endogeneous ETO gene activity. An evolutionary conserved region of 228 bp revealed potential cis-elements involved in transcription of ETO. Disruption of the evolutionary conserved GATA -636 consensus binding site repressed transactivation and disruption of the ETS1 -705 consensus binding site enhanced activity of the ETO promoter. The promoter was stimulated by overexpression of GATA-1 into erythroid/megakaryocytic cells. Electrophoretic mobility shift assay with erythroid/megakaryocytic cells showed specific binding of GATA-1 to the GATA -636 site. Furthermore, results from chromatin immunoprecipitation showed GATA-1 binding in vivo to the conserved region of the ETO promoter containing the -636 site. The results suggest that the GATA -636 site may have a role in activation of the ETO gene activity in cells with erythroid/megakaryocytic potential. Leukemia associated AML1-ETO strongly suppressed an ETO promoter reporter in erythroid/megakaryocytic cells.

CONCLUSIONS: We demonstrate that the GATA-1 transcription factor binds and transactivates the ETO proximal promoter in an erythroid/megakaryocytic-specific manner. Thus, trans-acting factors that are essential in erythroid/megakaryocytic differentiation govern ETO expression.

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keywords
Animals, Base Sequence, COS Cells, Cell Line, Tumor, Cercopithecus aethiops, Core Binding Factor Alpha 2 Subunit, GATA1 Transcription Factor, Gene Expression Regulation, HL-60 Cells, Humans, Leukemia, Erythroblastic, Acute, Leukemia, Myeloid, Acute, Megakaryocytes, Molecular Sequence Data, Oncogene Proteins, Fusion, Promoter Regions, Genetic, Proto-Oncogene Proteins, Transcription Factors, Journal Article, Research Support, Non-U.S. Gov't
in
BMC Molecular Biology
volume
11
article number
38
publisher
BioMed Central (BMC)
external identifiers
  • pmid:20487545
  • scopus:77952384773
ISSN
1471-2199
DOI
10.1186/1471-2199-11-38
language
English
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yes
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a9a5ca4f-8ec1-4b24-a0d8-6d4c9be67721
date added to LUP
2017-07-25 11:15:36
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2024-01-14 01:21:10
@article{a9a5ca4f-8ec1-4b24-a0d8-6d4c9be67721,
  abstract     = {{<p>BACKGROUND: The Eight-Twenty-One (ETO) nuclear co-repressor gene belongs to the ETO homologue family also containing Myeloid Translocation Gene on chromosome 16 (MTG16) and myeloid translocation Gene-Related protein 1 (MTGR1). By chromosomal translocations ETO and MTG16 become parts of fusion proteins characteristic of morphological variants of acute myeloid leukemia. Normal functions of ETO homologues have as yet not been examined. The goal of this work was to identify structural and functional promoter elements upstream of the coding sequence of the ETO gene in order to explore lineage-specific hematopoietic expression and get hints to function.</p><p>RESULTS: A putative proximal ETO promoter was identified within 411 bp upstream of the transcription start site. Strong ETO promoter activity was specifically observed upon transfection of a promoter reporter construct into erythroid/megakaryocytic cells, which have endogeneous ETO gene activity. An evolutionary conserved region of 228 bp revealed potential cis-elements involved in transcription of ETO. Disruption of the evolutionary conserved GATA -636 consensus binding site repressed transactivation and disruption of the ETS1 -705 consensus binding site enhanced activity of the ETO promoter. The promoter was stimulated by overexpression of GATA-1 into erythroid/megakaryocytic cells. Electrophoretic mobility shift assay with erythroid/megakaryocytic cells showed specific binding of GATA-1 to the GATA -636 site. Furthermore, results from chromatin immunoprecipitation showed GATA-1 binding in vivo to the conserved region of the ETO promoter containing the -636 site. The results suggest that the GATA -636 site may have a role in activation of the ETO gene activity in cells with erythroid/megakaryocytic potential. Leukemia associated AML1-ETO strongly suppressed an ETO promoter reporter in erythroid/megakaryocytic cells.</p><p>CONCLUSIONS: We demonstrate that the GATA-1 transcription factor binds and transactivates the ETO proximal promoter in an erythroid/megakaryocytic-specific manner. Thus, trans-acting factors that are essential in erythroid/megakaryocytic differentiation govern ETO expression.</p>}},
  author       = {{Ajore, Ram and Dhanda, Rakesh Singh and Gullberg, Urban and Olsson, Inge}},
  issn         = {{1471-2199}},
  keywords     = {{Animals; Base Sequence; COS Cells; Cell Line, Tumor; Cercopithecus aethiops; Core Binding Factor Alpha 2 Subunit; GATA1 Transcription Factor; Gene Expression Regulation; HL-60 Cells; Humans; Leukemia, Erythroblastic, Acute; Leukemia, Myeloid, Acute; Megakaryocytes; Molecular Sequence Data; Oncogene Proteins, Fusion; Promoter Regions, Genetic; Proto-Oncogene Proteins; Transcription Factors; Journal Article; Research Support, Non-U.S. Gov't}},
  language     = {{eng}},
  month        = {{05}},
  publisher    = {{BioMed Central (BMC)}},
  series       = {{BMC Molecular Biology}},
  title        = {{The leukemia associated ETO nuclear repressor gene is regulated by the GATA-1 transcription factor in erythroid/megakaryocytic cells}},
  url          = {{http://dx.doi.org/10.1186/1471-2199-11-38}},
  doi          = {{10.1186/1471-2199-11-38}},
  volume       = {{11}},
  year         = {{2010}},
}