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Pdgfrb is a direct regulatory target of TGFβ signaling in atrioventricular cushion mesenchymal cells

Cushion formation is the initial step for the development of valvuloseptal structures in mammalian hearts. TGFβ signaling plays critical roles in multiple steps of cushion morphogenesis. We used a newly developed conditional immortal atrioventricular cushion mesenchymal cell line, tsA58-AVM, to iden...

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Detalles Bibliográficos
Autores principales: Peng, Yin, Yan, Shun, Chen, Dongquan, Cui, Xiangqin, Jiao, Kai
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5398542/
https://www.ncbi.nlm.nih.gov/pubmed/28426709
http://dx.doi.org/10.1371/journal.pone.0175791
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author Peng, Yin
Yan, Shun
Chen, Dongquan
Cui, Xiangqin
Jiao, Kai
author_facet Peng, Yin
Yan, Shun
Chen, Dongquan
Cui, Xiangqin
Jiao, Kai
author_sort Peng, Yin
collection PubMed
description Cushion formation is the initial step for the development of valvuloseptal structures in mammalian hearts. TGFβ signaling plays critical roles in multiple steps of cushion morphogenesis. We used a newly developed conditional immortal atrioventricular cushion mesenchymal cell line, tsA58-AVM, to identify the TGFβ regulatory target genes through microarray analysis. Expression of ~1350 genes was significantly altered by TGFβ1 treatment. Subsequent bioinformatic analysis of TGFβ activated genes revealed that PDGF-BB signaling is the top hit as the potential upstream regulator. Among the 37 target molecules, 10 genes known to be involved in valve development and hemostasis were selected for quantitative reverse transcription polymerase chain reaction (qRT-PCR) analysis. Our results confirmed that they are all upregulated by TGFβ1 stimulation in tsA58-AVM cells and in primary atrioventricular cushion cells. We focused on examining regulation of Pdgfrb by TGFβ1, which encodes a tyrosine kinase receptor for PDGF-BB. We found that the ~150bp Pdgfrb promoter can respond to TGFβ stimulation and that this response relies on the two SP1 binding sites within the promoter. Co-immunoprecipitation analysis confirmed SP1 interacts with SMAD2 in a TGFβ-dependent fashion. Furthermore, SMAD2 is associated with the Pdgfrb promoter and this association is diminished by knocking down expression of Sp1. Our data therefore collectively suggest that upon TGFβ stimulation, SP1 recruits SMAD2 to the promoter of Pdgfrb to up-regulate its expression and thus Pdgfrb is a direct downstream target of the TGFβ/SMAD2 signaling.
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spelling pubmed-53985422017-05-04 Pdgfrb is a direct regulatory target of TGFβ signaling in atrioventricular cushion mesenchymal cells Peng, Yin Yan, Shun Chen, Dongquan Cui, Xiangqin Jiao, Kai PLoS One Research Article Cushion formation is the initial step for the development of valvuloseptal structures in mammalian hearts. TGFβ signaling plays critical roles in multiple steps of cushion morphogenesis. We used a newly developed conditional immortal atrioventricular cushion mesenchymal cell line, tsA58-AVM, to identify the TGFβ regulatory target genes through microarray analysis. Expression of ~1350 genes was significantly altered by TGFβ1 treatment. Subsequent bioinformatic analysis of TGFβ activated genes revealed that PDGF-BB signaling is the top hit as the potential upstream regulator. Among the 37 target molecules, 10 genes known to be involved in valve development and hemostasis were selected for quantitative reverse transcription polymerase chain reaction (qRT-PCR) analysis. Our results confirmed that they are all upregulated by TGFβ1 stimulation in tsA58-AVM cells and in primary atrioventricular cushion cells. We focused on examining regulation of Pdgfrb by TGFβ1, which encodes a tyrosine kinase receptor for PDGF-BB. We found that the ~150bp Pdgfrb promoter can respond to TGFβ stimulation and that this response relies on the two SP1 binding sites within the promoter. Co-immunoprecipitation analysis confirmed SP1 interacts with SMAD2 in a TGFβ-dependent fashion. Furthermore, SMAD2 is associated with the Pdgfrb promoter and this association is diminished by knocking down expression of Sp1. Our data therefore collectively suggest that upon TGFβ stimulation, SP1 recruits SMAD2 to the promoter of Pdgfrb to up-regulate its expression and thus Pdgfrb is a direct downstream target of the TGFβ/SMAD2 signaling. Public Library of Science 2017-04-20 /pmc/articles/PMC5398542/ /pubmed/28426709 http://dx.doi.org/10.1371/journal.pone.0175791 Text en © 2017 Peng 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
Peng, Yin
Yan, Shun
Chen, Dongquan
Cui, Xiangqin
Jiao, Kai
Pdgfrb is a direct regulatory target of TGFβ signaling in atrioventricular cushion mesenchymal cells
title Pdgfrb is a direct regulatory target of TGFβ signaling in atrioventricular cushion mesenchymal cells
title_full Pdgfrb is a direct regulatory target of TGFβ signaling in atrioventricular cushion mesenchymal cells
title_fullStr Pdgfrb is a direct regulatory target of TGFβ signaling in atrioventricular cushion mesenchymal cells
title_full_unstemmed Pdgfrb is a direct regulatory target of TGFβ signaling in atrioventricular cushion mesenchymal cells
title_short Pdgfrb is a direct regulatory target of TGFβ signaling in atrioventricular cushion mesenchymal cells
title_sort pdgfrb is a direct regulatory target of tgfβ signaling in atrioventricular cushion mesenchymal cells
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5398542/
https://www.ncbi.nlm.nih.gov/pubmed/28426709
http://dx.doi.org/10.1371/journal.pone.0175791
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