Cross-talk between regulators of myeloid development: C/EBPα binds and activates the promoter of the PU.1 gene
Abstract
CCAAT/enhancer-binding protein (C/EBP)alpha and PU.1 are required for myelopoiesis. Examination of the murine PU.1 promoter revealed several potential C/EBP-binding sites. Gel-shift assay demonstrated that C/EBPalpha expressed in 293T cells bound the site centered at -68 most potently. C/EBPalpha from 32D cl3 myeloid cell nuclear extracts also bound this site strongly, and endogenous C/EBPbeta did so to a lesser extent, whereas these C/EBP isoforms bound the neutrophil elastase promoter with equal affinity. The -68 site in the murine PU.1 promoter is conserved in the human PU.1 promoter. Mutation of the -68 C/EBP-binding site in a -85/+152 promoter segment linked to the luciferase cDNA reduced promoter activity fourfold in 293T cells in the presence of cotransfected C/EBPalpha and twofold in 32D cl3 myeloid cells. Induction of endogenous PU.1 RNA by C/EBPalpha-estradiol receptor (ER) in the presence of cycloheximide is obviated by mutation of the C/EBPalpha DNA-binding domain, and chromosomal immunoprecipitation demonstrated specific interaction of C/EBPalpha and C/EBPalpha-ER with the PU.1 promoter. Finally, PU.1 RNA is reduced several-fold in immortalized C/EBPalpha (-/-) compared with (+/-) cells. Together, these findings indicate that C/EBPalpha binds and activates the endogenous PU.1 gene in myeloid cells. Induction of PU.1 by C/EBPalpha may account for increased levels of PU.1 in myeloid as compared with B lymphoid cells and in this way, may contribute to the specificat…
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CEBPA binds to and induces Spi1 expression in HSPC.
“as CEBPA binds to and induces Spi1 expression in HSPC (Kummalue and Friedman, 2003; Yeamans et al., 2007).”
From PU.1 is required to restrain myelopoiesis during chronic inflammatory stress (2023) · cites this paper as Kummalue and Friedman, 2003
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References 40 · Cited by 52
- Authors
- Kummalue, T., & Friedman, A. D.
- Affiliations
- Johns Hopkins University (US)
- Corresponding author
- Friedman, Alan D
- ORCID
- Friedman, Alan D
- Venue
- Journal of Leukocyte Biology
- Publisher
- Oxford University Press (OUP)
- Published
- 2003-06-16
- Volume
- 74
- Issue
- 3
- Pages
- 464-470
- ISSN
- 0741-5400 (ISSN-L)
- 1938-3673
- OpenAlex
- W1996193200
- Cited by
- 52
- References
- 40
- Citations by year
- 2025: 1 · 2023: 1 · 2019: 3 · 2018: 2 · 2017: 2 · 2016: 1 · 2015: 3 · 2014: 2 · 2013: 3 · 2012: 3
- FWCI
- 1.03
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Journal of Leukocyte Biology (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Institutes of Health (HL62274)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Chromatin Dynamics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Retinoids in leukemia and cellular processes
- Health Sciences › Medicine › Hematology › Acute Myeloid Leukemia Research
- Keywords
- C/EBPα, PU.1, myeloid development, promoter activation, C/EBP binding site, C/EBPβ, estradiol receptor, myelopoiesis, transcriptional regulation
- MeSH terms
- Proto-Oncogene Protein Spi-1, Animals, Cell Nucleus, Cells, Cultured, Gene Expression Regulation, Luciferases, Promoter Regions, Genetic, Proto-Oncogene Proteins, Regulatory Sequences, Nucleic Acid, Transcription, Genetic, Transfection, Signal Transduction, Transcriptional Activation, Trans-Activators, Down-Regulation, Myeloid Cells, CCAAT-Enhancer-Binding Protein-alpha, Electrophoretic Mobility Shift Assay, Mice
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