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Regulation of early T-lineage gene expression and developmental progression by the progenitor cell transcription factor PU.1

Champhekar, Ameya and Damle, Sagar S. and Freedman, George and Carotta, Sebastian and Nutt, Stephen L. and Rothenberg, Ellen V. (2015) Regulation of early T-lineage gene expression and developmental progression by the progenitor cell transcription factor PU.1. Genes and Development, 29 (8). pp. 832-848. ISSN 0890-9369. PMCID PMC4403259. doi:10.1101/gad.259879.115. https://resolver.caltech.edu/CaltechAUTHORS:20150409-074847821

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Abstract

The ETS family transcription factor PU.1 is essential for the development of several blood lineages, including T cells, but its function in intrathymic T-cell precursors has been poorly defined. In the thymus, high PU.1 expression persists through multiple cell divisions in early stages but then falls sharply during T-cell lineage commitment. PU.1 silencing is critical for T-cell commitment, but it has remained unknown how PU.1 activities could contribute positively to T-cell development. Here we employed conditional knockout and modified antagonist PU.1 constructs to perturb PU.1 function stage-specifically in early T cells. We show that PU.1 is needed for full proliferation, restricting access to some non-T fates, and controlling the timing of T-cell developmental progression such that removal or antagonism of endogenous PU.1 allows precocious access to T-cell differentiation. Dominant-negative effects reveal that this repression by PU.1 is mediated indirectly. Genome-wide transcriptome analysis identifies novel targets of PU.1 positive and negative regulation affecting progenitor cell signaling and cell biology and indicating distinct regulatory effects on different subsets of progenitor cell transcription factors. Thus, in addition to supporting early T-cell proliferation, PU.1 regulates the timing of activation of the core T-lineage developmental program.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1101/gad.259879.115DOIArticle
http://genesdev.cshlp.org/content/29/8/832/suppl/DC1PublisherSupplemental Material
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4403259/PubMed CentralArticle
ORCID:
AuthorORCID
Rothenberg, Ellen V.0000-0002-3901-347X
Additional Information:© 2015 Champhekar et al.; Published by Cold Spring Harbor Laboratory Press. This article is distributed exclusively by Cold Spring Harbor Laboratory Press for the first six months after the full-issue publication date (see http://genesdev.cshlp.org/site/misc/terms.xhtml). After six months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/. Received February 5, 2015; revised version accepted March 17, 2015. Published in Advance April 6, 2015. We thank Sanket Acharya and Mark Zarnegar for the PU.1-Eng construct; Arthur Skoultchi for related PU.1 constructs used in preliminary studies; Hao Yuan Kueh for discussion on PCA analysis; Jonas Ungerbäck for sharing unpublished data; Robert Butler and Maria Quiloan for technical assistance; Rochelle Diamond, Diana Perez, and Pat Koen at the Caltech Flow Cytometry and Cell Sorting Facility for cell sorting; Scott Washburn and Ingrid Soto for animal care; and members of E.V.R.’s laboratory for helpful discussions. This work was supported by National Institutes of Health grants (CA90233, AI95943, and HD076915), the Louis A. Garfinkle Memorial Laboratory Fund, the Al Sherman Foundation, and the Albert Billings Ruddock Professorship to E.V. R., and Victorian State Government Operational Infrastructure Support, National Health and Medical Research Council of Australia Independent Research Institute Infrastructure Support grant 361646, Program grant 105492, and Fellowship 1058238 to S.L.N.
Funders:
Funding AgencyGrant Number
NIHCA90233
NIHAI95943
NIHHD076915
Louis A. Garfinkle Memorial Laboratory FundUNSPECIFIED
Al Sherman FoundationUNSPECIFIED
Albert Billings Ruddock ProfessorshipUNSPECIFIED
Victorian State Government Operational Infrastructure SupportUNSPECIFIED
National Health and Medical Research Council (NHMRC)361646
National Health and Medical Research Council (NHMRC)105492
National Health and Medical Research Council (NHMRC)1058238
Subject Keywords:conditional knockout; hematopoiesis; in vitro development; Notch; obligate repressor; RNA-seq
Issue or Number:8
PubMed Central ID:PMC4403259
DOI:10.1101/gad.259879.115
Record Number:CaltechAUTHORS:20150409-074847821
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20150409-074847821
Official Citation:Regulation of early T-lineage gene expression and developmental progression by the progenitor cell transcription factor PU.1 Ameya Champhekar, Sagar S. Damle, George Freedman, Sebastian Carotta, Stephen L. Nutt, and Ellen V. Rothenberg Genes Dev. April 15, 2015 29: 832-848; Published in Advance April 6, 2015, doi:10.1101/gad.259879.115
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:56504
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:09 Apr 2015 19:23
Last Modified:10 Nov 2021 21:00

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