Please use this identifier to cite or link to this item: https://doi.org/10.1371/journal.pone.0004268
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dc.titleGraded Smad2/3 activation is converted directly into levels of target gene expression in embryonic stem cells
dc.contributor.authorGuzman-Ayala M.
dc.contributor.authorLee K.L.
dc.contributor.authorMavrakis K.J.
dc.contributor.authorGoggolidou P.
dc.contributor.authorNorris D.P.
dc.contributor.authorEpiskopou V.
dc.date.accessioned2019-11-08T00:54:12Z
dc.date.available2019-11-08T00:54:12Z
dc.date.issued2009
dc.identifier.citationGuzman-Ayala M., Lee K.L., Mavrakis K.J., Goggolidou P., Norris D.P., Episkopou V. (2009). Graded Smad2/3 activation is converted directly into levels of target gene expression in embryonic stem cells. PLoS ONE 4 (1) : e4268. ScholarBank@NUS Repository. https://doi.org/10.1371/journal.pone.0004268
dc.identifier.issn19326203
dc.identifier.urihttps://scholarbank.nus.edu.sg/handle/10635/161843
dc.description.abstractThe Transforming Growth Factor (TGF) ? signalling family includes morphogens, such as Nodal and Activin, with important functions in vertebrate development. The concentration of the morphogen is critical for fate decisions in the responding cells. Smad2 and Smad3 are effectors of the Nodal/Activin branch of TGF? signalling: they are activated by receptors, enter the nucleus and directly transcribe target genes. However, there have been no studies correlating levels of Smad2/3 activation with expression patterns of endogenous target genes in a developmental context over time. We used mouse Embryonic Stem (ES) cells to create a system whereby levels of activated Smad2/3 can be manipulated by an inducible constitutively active receptor (Alk4*) and an inhibitor (SB-431542) that blocks specifically Smad2/3 activation. The transcriptional responses were analysed by microarrays at different time points during activation and repression. We identified several genes that follow faithfully and reproducibly the Smad2/3 activation profile. Twenty-seven of these were novel and expressed in the early embryo downstream of Smad2/3 signalling. As they responded to Smad2/3 activation in the absence of protein synthesis, they were considered direct. These immediate responsive genes included negative intracellular feedback factors, like SnoN and I-Smad7, which inhibit the transcriptional activity of Smad2/3. However, their activation did not lead to subsequent repression of target genes over time, suggesting that this type of feedback is inefficient in ES cells or it is counteracted by mechanisms such as ubiquitin-mediated degradation by Arkadia. Here we present an ES cell system along with a database containing the expression profile of thousands of genes downstream of Smad2/3 activation patterns, in the presence or absence of protein synthesis. Furthermore, we identify primary target genes that follow proportionately and with high sensitivity changes in Smad2/3 levels over 15-30 hours. The above system and resource provide tools to study morphogen function in development. � 2009 Guzman-Ayala et al.
dc.rightsAttribution 4.0 International
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/
dc.sourceUnpaywall 20191101
dc.subject4 [4 (1,3 benzodioxol 5 yl) 5 (2 pyridinyl) 1h imidazol 2 yl]benzamide
dc.subjectSmad2 protein
dc.subjectSmad3 protein
dc.subjectSmad7 protein
dc.subject1,3 dioxolane derivative
dc.subject4 (5 benzo(1,3)dioxol 5 yl 4 pyridin 2 yl 1H imidazol 2 yl)benzamide
dc.subject4-(5-benzo(1,3)dioxol-5-yl-4-pyridin-2-yl-1H-imidazol-2-yl)benzamide
dc.subjectactivin receptor 1
dc.subjectAcvr1b protein, mouse
dc.subjectArkadia protein, mouse
dc.subjectbenzamide derivative
dc.subjectNodal protein, mouse
dc.subjectprotein Nodal
dc.subjectSmad2 protein
dc.subjectSmad2 protein, mouse
dc.subjectSmad3 protein
dc.subjectSmad3 protein, mouse
dc.subjecttransforming growth factor beta
dc.subjectubiquitin
dc.subjectanimal cell
dc.subjectanimal tissue
dc.subjectarticle
dc.subjectcontrolled study
dc.subjectDNA microarray
dc.subjectembryo
dc.subjectembryo development
dc.subjectembryonic stem cell
dc.subjectfeedback system
dc.subjectgene activation
dc.subjectgene expression profiling
dc.subjectgene identification
dc.subjectgene repression
dc.subjectgenetic transcription
dc.subjectmorphogenesis
dc.subjectmouse
dc.subjectnonhuman
dc.subjectnucleotide sequence
dc.subjectprotein synthesis
dc.subjectsignal transduction
dc.subjecttranscription regulation
dc.subjectunindexed sequence
dc.subjectanimal
dc.subjectbiological model
dc.subjectflow cytometry
dc.subjectgene expression regulation
dc.subjectmetabolism
dc.subjectVertebrata
dc.subjectActivin Receptors, Type I
dc.subjectAnimals
dc.subjectBenzamides
dc.subjectDioxoles
dc.subjectEmbryonic Stem Cells
dc.subjectFlow Cytometry
dc.subjectGene Expression Regulation, Developmental
dc.subjectMice
dc.subjectModels, Biological
dc.subjectNodal Protein
dc.subjectSignal Transduction
dc.subjectSmad2 Protein
dc.subjectSmad3 Protein
dc.subjectTranscription, Genetic
dc.subjectTransforming Growth Factor beta
dc.subjectUbiquitin
dc.typeArticle
dc.contributor.departmentDUKE-NUS MEDICAL SCHOOL
dc.description.doi10.1371/journal.pone.0004268
dc.description.sourcetitlePLoS ONE
dc.description.volume4
dc.description.issue1
dc.description.pagee4268
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