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C/EBPα induces Ebf1 gene expression in common lymphoid progenitors

C/EBPα is required for formation of granulocyte-monocyte progenitors (GMP) and also participates in B lymphopoiesis. The common lymphoid progenitor (CLP) and preproB populations but not proB cells express Cebpa, and pan-hematopoietic deletion of the +37 kb Cebpa enhancer using Mx1-Cre leads not only...

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Autores principales: Barberi, Theresa, Cui, Cheng, Friedman, Alan D.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7746190/
https://www.ncbi.nlm.nih.gov/pubmed/33332417
http://dx.doi.org/10.1371/journal.pone.0244161
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author Barberi, Theresa
Cui, Cheng
Friedman, Alan D.
author_facet Barberi, Theresa
Cui, Cheng
Friedman, Alan D.
author_sort Barberi, Theresa
collection PubMed
description C/EBPα is required for formation of granulocyte-monocyte progenitors (GMP) and also participates in B lymphopoiesis. The common lymphoid progenitor (CLP) and preproB populations but not proB cells express Cebpa, and pan-hematopoietic deletion of the +37 kb Cebpa enhancer using Mx1-Cre leads not only to reduced GMP but also to 2-fold reduced marrow preproB and >15-fold reduced proB and preB cells. We now show that IL7Rα-Cre-mediated deletion of the +37 kb Cebpa enhancer, which occurs in 89% of Ly6D(+) and 65% of upstream Ly6D(-) CLP, leads to a 2-fold reduction of both preproB and proB cells, and a 3-fold reduction in preB cells, with no impact on GMP numbers. These data support a direct role for C/EBPα during B lineage development, with reduced enhancer deletion in Ly6D(-) CLP mediated by IL7Rα-Cre diminishing the effect on B lymphopoiesis compared to that seen with Mx1-Cre. Amongst mRNAs encoding key transcriptional regulators that initiate B lymphoid specification (PU.1, E2A, IKAROS, EBF1, FOXO1, and BACH2), only Ebf1 levels are altered in CLP upon Mx1-Cre-mediated Cebpa enhancer deletion, with Ebf1 reduced ~40-fold in Flt3(+)Sca-1(int)c-kit(int)IL7Rα(+) CLP. In addition, Cebpa and Ebf1 RNAs were 4- and 14-fold higher in hCD4(+) versus hCD4(-) CLP from Cebpa-hCD4 transgenic mice. Histone modification ChIP-Seq data for CLP indicate the presence of active, intronic Ebf1 enhancers located 270 and 280 kb upstream of the transcription start sites. We identified a cis element in this region that strongly binds C/EBPα using the electrophoretic mobility shift assay. Mutation of this C/EBPα-binding site in an Ebf1 enhancer-TK-luciferase reporter leads to a 4-fold reduction in C/EBPα-mediated trans-activation. These findings support a model of B lymphopoiesis in which induction of Ebf1 by C/EBPα in a subset of CLP contributes to initiation of B lymphopoiesis.
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spelling pubmed-77461902020-12-31 C/EBPα induces Ebf1 gene expression in common lymphoid progenitors Barberi, Theresa Cui, Cheng Friedman, Alan D. PLoS One Research Article C/EBPα is required for formation of granulocyte-monocyte progenitors (GMP) and also participates in B lymphopoiesis. The common lymphoid progenitor (CLP) and preproB populations but not proB cells express Cebpa, and pan-hematopoietic deletion of the +37 kb Cebpa enhancer using Mx1-Cre leads not only to reduced GMP but also to 2-fold reduced marrow preproB and >15-fold reduced proB and preB cells. We now show that IL7Rα-Cre-mediated deletion of the +37 kb Cebpa enhancer, which occurs in 89% of Ly6D(+) and 65% of upstream Ly6D(-) CLP, leads to a 2-fold reduction of both preproB and proB cells, and a 3-fold reduction in preB cells, with no impact on GMP numbers. These data support a direct role for C/EBPα during B lineage development, with reduced enhancer deletion in Ly6D(-) CLP mediated by IL7Rα-Cre diminishing the effect on B lymphopoiesis compared to that seen with Mx1-Cre. Amongst mRNAs encoding key transcriptional regulators that initiate B lymphoid specification (PU.1, E2A, IKAROS, EBF1, FOXO1, and BACH2), only Ebf1 levels are altered in CLP upon Mx1-Cre-mediated Cebpa enhancer deletion, with Ebf1 reduced ~40-fold in Flt3(+)Sca-1(int)c-kit(int)IL7Rα(+) CLP. In addition, Cebpa and Ebf1 RNAs were 4- and 14-fold higher in hCD4(+) versus hCD4(-) CLP from Cebpa-hCD4 transgenic mice. Histone modification ChIP-Seq data for CLP indicate the presence of active, intronic Ebf1 enhancers located 270 and 280 kb upstream of the transcription start sites. We identified a cis element in this region that strongly binds C/EBPα using the electrophoretic mobility shift assay. Mutation of this C/EBPα-binding site in an Ebf1 enhancer-TK-luciferase reporter leads to a 4-fold reduction in C/EBPα-mediated trans-activation. These findings support a model of B lymphopoiesis in which induction of Ebf1 by C/EBPα in a subset of CLP contributes to initiation of B lymphopoiesis. Public Library of Science 2020-12-17 /pmc/articles/PMC7746190/ /pubmed/33332417 http://dx.doi.org/10.1371/journal.pone.0244161 Text en © 2020 Barberi et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Barberi, Theresa
Cui, Cheng
Friedman, Alan D.
C/EBPα induces Ebf1 gene expression in common lymphoid progenitors
title C/EBPα induces Ebf1 gene expression in common lymphoid progenitors
title_full C/EBPα induces Ebf1 gene expression in common lymphoid progenitors
title_fullStr C/EBPα induces Ebf1 gene expression in common lymphoid progenitors
title_full_unstemmed C/EBPα induces Ebf1 gene expression in common lymphoid progenitors
title_short C/EBPα induces Ebf1 gene expression in common lymphoid progenitors
title_sort c/ebpα induces ebf1 gene expression in common lymphoid progenitors
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7746190/
https://www.ncbi.nlm.nih.gov/pubmed/33332417
http://dx.doi.org/10.1371/journal.pone.0244161
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