Core Binding Factor Cannot Synergistically Activate the Myeloperoxidase Proximal Enhancer in Immature Myeloid Cells without c-Myb

Britos-Bray, Martin; Friedman, Alan D.

doi:10.1128/mcb.17.9.5127

Cited by 94Genomics and Chromatin DynamicsAcute Myeloid Leukemia ResearchRetinoids in leukemia and cellular processes

Core Binding Factor Cannot Synergistically Activate the Myeloperoxidase Proximal Enhancer in Immature Myeloid Cells without c-Myb

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Britos-Bray, Martin, Friedman, Alan D.·1997·Molecular and Cellular Biology

DOI 10.1128/mcb.17.9.5127PMC PMC232363

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Abstract

The myeloperoxidase (MPO) gene is transcribed specifically in immature myeloid cells and is regulated in part by a 414-bp proximal enhancer. Mutation of a core binding factor (CBF)-binding site at -288 decreased enhancer activity 30-fold in 32D cl3 myeloid cells cultured in granulocyte colony-stimulating factor (G-CSF). A novel functional analysis, linking the CBF-binding site to an enhancer deletion series, located at -147 an evolutionarily conserved c-Myb-binding site which was required for optimal enhancer activity and synergy with CBF in 32D cells. These sites cooperated in isolation and independent of a precise spacing. Deletional analysis carried out in the absence of the c-Myb-binding site at -147 located at -301 a second c-Myb-binding site which also synergized with CBF to activate the enhancer. A GA-rich region at -162 contributed to cooperation with CBF when the adjacent c-Myb-binding site was intact. Mutation of both c-Myb-binding sites in the context of the entire enhancer greatly impaired activation by endogenous CBF in 32D cells. Similarly, activation by c-Myb was impaired in constructs lacking the CBF-binding site. CBF and c-Myb were required for induction of MPO proximal enhancer activity when 32D cells differentiated in response to G-CSF. A fusion protein containing the Gal4 DNA-binding domain and the AML-1B activation domain, amino acids 216 to 480, activated transcription alone and cooperatively with c-Myb in nonmyeloid CV-1 cells. Determining how CBF and c-Myb synergize in myeloid cells might contribute to our understanding of leukemogenesis by the AML1-ETO, AML1-MDS1, CBFbeta-SMMHC, and v-Myb oncoproteins.

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The region from amino acids 1 to 317 is important for synergy between AML1 and c-Myb.
“a region important for synergy between AML1 and c-Myb (6)”
From Multiple Functional Domains of AML1: PU.1 and C/EBPα Synergize with Different Regions of AML1 (1998) · cites this paper as [6]
The Discussion (page unavailable) identifies AML-1B regions 216–289 and 278–480 as required for optimal cooperation with c-Myb, while saying its DNA-binding domain is dispensable. This supports the importance of part of amino acids 1–317, but not that the specified 1–317 region as a whole is the region required for synergy; the paper also identifies a required segment extending beyond residue 317. (LLM, conf=0.85)
Experimental — accuracy not yet measured
The myeloid-specific expression of neutrophil elastase, M-CSF receptor, and myeloperoxidase is mainly controlled through the concerted and synergistic action of RUNX1 with factors such as Myb, PU.1, and C/EBP, which bind closely located elements on their respective promoter regions.
“The myeloid-specific expression of the neutrophil elastase, 58 M-CSF receptor, 29,30 and myeloperoxidase 27 is mainly controlled through the concerted and synergistic action of RUNX1 with factors such as Myb, PU.1, and C/EBP, which bind closely located elements on their respective promoter regions.”
From RUNX/AML and C/EBP factors regulate CD11a integrin expression in myeloid cells through overlapping regulatory elements. (2003) · cites this paper as [27]

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Ford, A. M., C. A. Bennett, L. E. Healy, M. Towatari, M. F. Greaves, and T. Enver. 1996. Regulation of the myeloperoxidase enhancer binding proteins PU.1, C/EBP␣, ␤, and ␦ during granulocyte-lineage specification. Proc. Natl. Acad. Sci. USA 93:10838-10843.

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Authors: Britos-Bray, M., & Friedman, A. D.
Affiliations: Johns Hopkins University (US)
Corresponding author: Friedman, Alan D.
ORCID: Friedman, Alan D.
Venue: Molecular and Cellular Biology
Publisher: Informa UK Limited
Published: 1997-09
Volume: 17
Issue: 9
Pages: 5127-5135
ISSN: 0270-7306 (ISSN-L)
1098-5549
DOI: 10.1128/mcb.17.9.5127
PMC: PMC232363
OpenAlex: W2152668799
Cited by: 94
References: 47
Citations by year: 2017: 1 · 2016: 1 · 2015: 1 · 2014: 1 · 2012: 3
FWCI: 2.96
Type: journal article
Language: en
Open access: bronze
doi.org
Licence: journals.asm.org/non-commercial-tdm-license (tdm)
Locations: Molecular and Cellular Biology (publishedVersion)
PubMed Central (publishedVersion)
PubMed (publishedVersion)
Funders: National Institutes of Health
National Heart, Lung, and Blood Institute (R01 HL15388)
Topics: Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Chromatin Dynamics
Health Sciences › Medicine › Hematology › Acute Myeloid Leukemia Research
Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Retinoids in leukemia and cellular processes
Keywords: myeloperoxidase, core binding factor, MYB, transcriptional synergy, enhancer activation, immature myeloid cells, 32D cells, RUNX1, AML1-ETO, leukemogenesis, transcription factor binding sites
MeSH terms: Animals, Base Sequence, Binding Sites, Cell Differentiation, Cells, Cultured, DNA-Binding Proteins, Enhancer Elements, Genetic, Enzyme Activation, Hematopoietic Stem Cells, Molecular Sequence Data, Peroxidase, Neoplasm Proteins, Proto-Oncogene Proteins, Transcription Factors, Transcription, Genetic, Trans-Activators, Proto-Oncogene Proteins c-myb, Core Binding Factors, Mice

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Core Binding Factor Cannot Synergistically Activate the Myeloperoxidase Proximal Enhancer in Immature Myeloid Cells without c-Myb

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