Architecture of a lymphomyeloid developmental switch controlled by PU.1, Notch and Gata3
Open accessAbstract
Hematopoiesis is a classic system with which to study developmental potentials and to investigate gene regulatory networks that control choices among alternate lineages. T-cell progenitors seeding the thymus retain several lineage potentials. The transcription factor PU.1 is involved in the decision to become a T cell or a myeloid cell, and the developmental outcome of expressing PU.1 is dependent on exposure to Notch signaling. PU.1-expressing T-cell progenitors without Notch signaling often adopt a myeloid program, whereas those exposed to Notch signals remain in a T-lineage pathway. Here, we show that Notch signaling does not alter PU.1 transcriptional activity by degradation/alteration of PU.1 protein. Instead, Notch signaling protects against the downregulation of T-cell factors so that a T-cell transcriptional network is maintained. Using an early T-cell line, we describe two branches of this network. The first involves inhibition of E-proteins by PU.1 and the resulting inhibition of Notch signaling target genes. Effects of E-protein inhibition can be reversed by exposure to Notch signaling. The second network is dependent on the ability of PU.1 to inhibit important T-cell transcription factor genes such as Myb, Tcf7 and Gata3 in the absence of Notch signaling. We show that maintenance of Gata3 protein levels by Myb and Notch signaling is linked to the ability to retain T-cell identity in response to PU.1.
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Even artificially high-level PU.1 can redirect differentiation to myeloid or dendritic fates only if Notch signaling is reduced.
“Even artificially high-level PU.1 could only redirect the differentiation of the cells to myeloid or dendritic fates if Notch signaling were reduced (49, 50, 58).”
From Mechanisms of Action of Hematopoietic Transcription Factor PU.1 in Initiation of T-Cell Development (2019) · cites this paper as [58]
The section ‘Pro-T cells with high levels of PU.1 protein are able to resist Mac1 upregulation in the presence of Notch signaling’ (page not provided) says Notch reduces the proportion of cells showing myeloid diversion, but Mac1 could still be induced at the highest PU.1 levels while Notch ligand was present. Thus, reduced Notch signaling was not an absolute requirement, contrary to ‘only if.’ (LLM, conf=0.87)
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Experimental — accuracy not yet measuredEven with PU.1 transduction, Scid.adh.2C2 cells do not restore the full chromatin accessibility landscape of ETP and DN2a pro-T cells and reactivate little of the normal context program.
“Even with PU.1 transduction, Scid.adh.2C2 cells do not restore the full chromatin accessibility landscape of ETP and DN2a stage pro-T cells, and despite detectable upregulation of a few other early pro-T cell genes (e.g., Bcl11a and Lyl1), the transduced cells as a whole reactivate little of the program that forms the normal context for endogenous PU.1 activity in pro-T cells (58, 79, 85).”
From Mechanisms of Action of Hematopoietic Transcription Factor PU.1 in Initiation of T-Cell Development (2019) · cites this paper as [58]
The section on the Scid.adh.2C2 cell line describes it as DN3-like and reports that PU.1 transduction upregulated Mac1 in only a fraction of cells. The paper also reports PU.1-dependent repression of T-cell genes, which is consistent with an incomplete early pro-T-cell response. But our copy does not compare these cells’ chromatin accessibility with ETP or DN2a cells, or establish how much of the normal early pro-T-cell program they reactivate. (LLM, conf=0.74)
Experimental — accuracy not yet measuredInstead of re-acquiring a progenitor-like state, cells appear to trans-differentiate to a dendritic-cell or macrophage developmental program.
“Instead of re-acquiring aspects of a progenitor-like state, the cells appear to trans-differentiate to a dendritic-cell or macrophage developmental program (48, 50, 58).”
From Mechanisms of Action of Hematopoietic Transcription Factor PU.1 in Initiation of T-Cell Development (2019) · cites this paper as [58]
The Results section on diversion in Scid.adh.2C2 cells (page unavailable) reports that PU.1-diverted cells downregulated T-cell genes and upregulated macrophage-associated genes, supporting a shift toward a macrophage program. The passages do not establish diversion to a dendritic-cell program or show that the cells bypass re-acquisition of progenitor-like features. (LLM, conf=0.83)
Experimental — accuracy not yet measuredScid.adh.2C2 cells respond in an all-or-none way to forced PU.1, coordinately upregulating myeloid/dendritic genes and downregulating T-cell genes in a discrete fraction increasing with PU.1 levels.
“They respond in an all-or-none way to forced expression of PU.1, coordinately upregulating myeloid- or dendritic-cell associated genes and downregulating T-cell genes in a discrete fraction of the cells that increases with increasing levels of PU.1 (47, 58)”
From Mechanisms of Action of Hematopoietic Transcription Factor PU.1 in Initiation of T-Cell Development (2019) · cites this paper as [58]
The section on Scid.adh.2C2 cells reports an all-or-none response to forced PU.1, with Mac1 upregulated in a fraction of cells; the gene-expression sections link diversion to repression of T-cell genes and induction of myeloid markers (page unavailable in our copy). However, the passage relating PU.1 protein levels to the fraction of Mac1-positive cells concerns fetal thymocytes, not Scid.adh.2C2 cells. This source therefore does not establish that the responding fraction of Scid.adh.2C2 cells increases with PU.1 level. (LLM, conf=0.85)
Experimental — accuracy not yet measuredHigh-dose PU.1 can inhibit expression of multiple Notch target genes and Notch1 itself.
“First, as noted above, high-dose PU.1 can inhibit expression of multiple Notch target genes and Notch1 itself (58, 85).”
From Mechanisms of Action of Hematopoietic Transcription Factor PU.1 in Initiation of T-Cell Development (2019) · cites this paper as [58]
The Results (RESEARCH ARTICLE, Fig. 2B–D; page unavailable) report that PU.1 inhibits several Notch target genes, including Nrarp, HEBalt and Hes1. The passages found do not report that PU.1 inhibits expression of Notch1 itself; the mention of Notch1 in the Dissection section concerns experimentally introduced active Notch1. (LLM, conf=0.85)
Experimental — accuracy not yet measured
PU.1 provides positive input into a discrete subset of Phase 1 regulatory genes, with Mef2c, Lmo2, Bcl11a, and often also Hhex responding over a range of different tests.
“Varied PU.1 gain and loss of function perturbations in the DN2-DN3 stages show that PU.1 does provide positive input into a discrete subset of Phase 1 regulatory genes, with Mef2c, Lmo2, Bcl11a, and often also Hhex responding over a range of different tests (58, 79, 80, 85).”
From Mechanisms of Action of Hematopoietic Transcription Factor PU.1 in Initiation of T-Cell Development (2019) · cites this paper as [58]
In the section on fetal-thymocyte responses to high-level PU.1 (page not supplied), the paper reports PU.1-dependent upregulation of Bcl11a and Hhex and effects on Lmo2 in DN2 cells; it says the Hhex and Lmo2 effects were not significant in DN3 cells. The cited copy does not show Mef2c responding or establish the claimed pattern across both gain- and loss-of-function tests. (LLM, conf=0.86)
Experimental — accuracy not yet measuredPU.1 actions are Notch-sensitive.
“PU.1 actions are Notch-sensitive.”
From Mechanisms of Action of Hematopoietic Transcription Factor PU.1 in Initiation of T-Cell Development (2019) · cites this paper as [58]
In primary fetal-derived pro-T cells and a DN3-like cell line, genes affected by a fixed PU.1 level depended strongly on the strength of Notch signaling.
“In primary fetal-derived pro-T cells and in a DN3-like cell line, the particular genes affected by a given, fixed level of PU.1 in the cells depended strongly on the strength of Notch signaling being induced in the cells at the time (58).”
From Mechanisms of Action of Hematopoietic Transcription Factor PU.1 in Initiation of T-Cell Development (2019) · cites this paper as [58]
In primary pro-T cells and Scid.adh.2C2 cells, highly overexpressed PU.1 appears to inhibit Notch signaling, as measured by downregulation of Notch target genes and Notch1 itself.
“In primary pro-T cells and Scid.adh.2C2 cells, highly overexpressed PU.1 appears to inhibit Notch signaling, as measured by downregulation of Notch target genes and even Notch1 itself.”
From Mechanisms of Action of Hematopoietic Transcription Factor PU.1 in Initiation of T-Cell Development (2019) · cites this paper as [58]
The gene network switch underlies stochastic, switch-like behavior of individual pro-T or Scid.adh.2C2 cells when forced to express high-level PU.1.
“this gene network switch underlies the stochastic, switch-like behavior of individual pro-T or Scid.adh.2C2 cells when forced to express high-level PU.1 (47, 58).”
From Mechanisms of Action of Hematopoietic Transcription Factor PU.1 in Initiation of T-Cell Development (2019) · cites this paper as [58]
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References 42 · Cited by 95
Research works printed but not linked: 2
- Germar, K., Dose, M., Konstantinou, T., Zhang, J., Wang, H., Lobry, C., Arnett, K. L., Blacklow, S. C., Aifantis, I., Aster, J. C. et al. (2011). T-cell factor 1 is a gatekeeper for T-cell specification in response to Notch signaling. Proc. Natl. Acad. Sci. USA 108, 20060-20065.
- Schwartz, R., Engel, I., Fallahi-Sichani, M., Petrie, H. T. and Murre, C. (2006). Gene expression patterns define novel roles for E47 in cell cycle progression, cytokine-mediated signaling, and T lineage development. Proc. Natl. Acad. Sci. USA 103, 9976-9981.
- Authors
- Del Real, M. M., & Rothenberg, E. V.
- Affiliations
- Division of Biology 156-29, California Institute of Technology, Pasadena, CA 91125, USA.
- California Institute of Technology (US)
- Corresponding author
- Rothenberg, Ellen V.
- ORCID
- Rothenberg, Ellen V.
- Venue
- Development
- Publisher
- The Company of Biologists
- Published
- 2013-03-15
- Volume
- 140
- Issue
- 6
- Pages
- 1207-1219
- ISSN
- 0950-1991 (ISSN-L)
- 1477-9129
- PMC
- PMC3585658
- OpenAlex
- W2033544609
- Cited by
- 95
- References
- 42
- Citations by year
- 2026: 2 · 2025: 4 · 2024: 6 · 2023: 9 · 2022: 5 · 2021: 7 · 2020: 8 · 2019: 14 · 2018: 8 · 2017: 7 · 2016: 5 · 2015: 9 · 2014: 6 · 2013: 5
- FWCI
- 3.74
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Development (publishedVersion)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- CaltechAUTHORS (California Institute of Technology) (submittedVersion)
- Funders
- National Institutes of Health
- National Cancer Institute (R01CA090233, CA90233, CA90233-08S1)
- National Institute of General Medical Sciences (T32GM-07616, T32GM007616)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Developmental Biology and Gene Regulation
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Chromatin Dynamics
- Life Sciences › Immunology and Microbiology › Immunology › T-cell and B-cell Immunology
- Keywords
- PU.1, Notch signaling, GATA3, T cell lineage commitment, myeloid lineage commitment, T cell progenitors, E protein, MYB, TCF1, transcriptional regulatory networks, lineage choice, hematopoiesis
- MeSH terms
- Proto-Oncogene Protein Spi-1, Animals, Cell Differentiation, Cells, Cultured, Embryo, Mammalian, Genes, Switch, Hematopoiesis, Mice, Inbred C57BL, Mice, Transgenic, Models, Biological, Proto-Oncogene Proteins, T-Lymphocytes, Signal Transduction, Trans-Activators, Gene Expression Regulation, Developmental, Cell Lineage, Proto-Oncogene Proteins c-myb, Myeloid Progenitor Cells, GATA3 Transcription Factor, Mice, Receptors, Notch, Thymocytes
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