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https://doi.org/10.1371/journal.pone.0004268
DC Field | Value | |
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dc.title | Graded Smad2/3 activation is converted directly into levels of target gene expression in embryonic stem cells | |
dc.contributor.author | Guzman-Ayala M. | |
dc.contributor.author | Lee K.L. | |
dc.contributor.author | Mavrakis K.J. | |
dc.contributor.author | Goggolidou P. | |
dc.contributor.author | Norris D.P. | |
dc.contributor.author | Episkopou V. | |
dc.date.accessioned | 2019-11-08T00:54:12Z | |
dc.date.available | 2019-11-08T00:54:12Z | |
dc.date.issued | 2009 | |
dc.identifier.citation | Guzman-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.issn | 19326203 | |
dc.identifier.uri | https://scholarbank.nus.edu.sg/handle/10635/161843 | |
dc.description.abstract | The 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.rights | Attribution 4.0 International | |
dc.rights.uri | http://creativecommons.org/licenses/by/4.0/ | |
dc.source | Unpaywall 20191101 | |
dc.subject | 4 [4 (1,3 benzodioxol 5 yl) 5 (2 pyridinyl) 1h imidazol 2 yl]benzamide | |
dc.subject | Smad2 protein | |
dc.subject | Smad3 protein | |
dc.subject | Smad7 protein | |
dc.subject | 1,3 dioxolane derivative | |
dc.subject | 4 (5 benzo(1,3)dioxol 5 yl 4 pyridin 2 yl 1H imidazol 2 yl)benzamide | |
dc.subject | 4-(5-benzo(1,3)dioxol-5-yl-4-pyridin-2-yl-1H-imidazol-2-yl)benzamide | |
dc.subject | activin receptor 1 | |
dc.subject | Acvr1b protein, mouse | |
dc.subject | Arkadia protein, mouse | |
dc.subject | benzamide derivative | |
dc.subject | Nodal protein, mouse | |
dc.subject | protein Nodal | |
dc.subject | Smad2 protein | |
dc.subject | Smad2 protein, mouse | |
dc.subject | Smad3 protein | |
dc.subject | Smad3 protein, mouse | |
dc.subject | transforming growth factor beta | |
dc.subject | ubiquitin | |
dc.subject | animal cell | |
dc.subject | animal tissue | |
dc.subject | article | |
dc.subject | controlled study | |
dc.subject | DNA microarray | |
dc.subject | embryo | |
dc.subject | embryo development | |
dc.subject | embryonic stem cell | |
dc.subject | feedback system | |
dc.subject | gene activation | |
dc.subject | gene expression profiling | |
dc.subject | gene identification | |
dc.subject | gene repression | |
dc.subject | genetic transcription | |
dc.subject | morphogenesis | |
dc.subject | mouse | |
dc.subject | nonhuman | |
dc.subject | nucleotide sequence | |
dc.subject | protein synthesis | |
dc.subject | signal transduction | |
dc.subject | transcription regulation | |
dc.subject | unindexed sequence | |
dc.subject | animal | |
dc.subject | biological model | |
dc.subject | flow cytometry | |
dc.subject | gene expression regulation | |
dc.subject | metabolism | |
dc.subject | Vertebrata | |
dc.subject | Activin Receptors, Type I | |
dc.subject | Animals | |
dc.subject | Benzamides | |
dc.subject | Dioxoles | |
dc.subject | Embryonic Stem Cells | |
dc.subject | Flow Cytometry | |
dc.subject | Gene Expression Regulation, Developmental | |
dc.subject | Mice | |
dc.subject | Models, Biological | |
dc.subject | Nodal Protein | |
dc.subject | Signal Transduction | |
dc.subject | Smad2 Protein | |
dc.subject | Smad3 Protein | |
dc.subject | Transcription, Genetic | |
dc.subject | Transforming Growth Factor beta | |
dc.subject | Ubiquitin | |
dc.type | Article | |
dc.contributor.department | DUKE-NUS MEDICAL SCHOOL | |
dc.description.doi | 10.1371/journal.pone.0004268 | |
dc.description.sourcetitle | PLoS ONE | |
dc.description.volume | 4 | |
dc.description.issue | 1 | |
dc.description.page | e4268 | |
Appears in Collections: | Staff Publications Elements |
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