scholarly journals Genetically regulated humanNODALsplice variants are differentially post-transcriptionally processed and functionally distinct

2018 ◽  
Author(s):  
Scott D Findlay ◽  
Olena Bilyk ◽  
Kiefer Lypka ◽  
Andrew J Waskiewicz ◽  
Lynne-Marie Postovit
Keyword(s):  
Transcriptional Regulation ◽  
Embryonic Stem ◽  
Proteolytic Processing ◽  
Model Organisms ◽  
Functional Capability ◽  
Selective Depletion ◽  
Enhanced Secretion ◽  
Nodal Signal ◽  
Post Transcriptional Regulation ◽  
Human Specific

AbstractNODALis a morphogen essential for early embryonic development in vertebrates. Since much of our understanding ofNODALcomes from model organisms, we aimed to directly assess post-transcriptional regulation of human NODAL with specific attention to a newly discovered human-specific NODAL splice variant. Selective depletion of the NODAL variant in human embryonic stem cells resulted in increased LIFR levels, while total NODAL knockdown resulted in a decrease of several markers of pluripotency. The NODAL variant did not transmit a canonical NODAL signal in zebrafish embryos, but may share some functional capability with canonical NODAL in cancer cells. At the protein level, disruption of disulfide bond formation dramatically enhanced proteolytic processing of NODAL. Disruption of NODAL N-glycosylation decreased its secretion but not extracellular stability, and a novel N-glycosylation in the NODAL variant contributed to enhanced secretion. Collectively, this work offers a direct and precise account of post-transcriptional regulation of human NODAL.

scholarly journals Quantitative Proteomics Analysis Demonstrates Post-transcriptional Regulation of Embryonic Stem Cell Differentiation to Hematopoiesis

2007 ◽  
Vol 7 (3) ◽  
pp. 459-472 ◽  
Author(s):  
Andrew J. K. Williamson ◽  
Duncan L. Smith ◽  
David Blinco ◽  
Richard D. Unwin ◽  
Stella Pearson ◽  
...  
Keyword(s):  
Transcriptional Regulation ◽  
Stem Cell ◽  
Cell Differentiation ◽  
Embryonic Stem Cell ◽  
Quantitative Proteomics ◽  
Embryonic Stem ◽  
Stem Cell Differentiation ◽  
Embryonic Stem Cell Differentiation ◽  
Proteomics Analysis ◽  
Post Transcriptional Regulation

Essential Cytoplasmic Role of the Histone Lysine Methyltransferase Setdb1 in Post-Transcriptional Regulation in Embryonic Stem Cells

2021 ◽  
Author(s):  
Roberta Rapone ◽  
Laurence Del Maestro ◽  
Costas Bouyioukos ◽  
Sonia Albini ◽  
Paola Cruz-Tapias ◽  
...  
Keyword(s):  
Gene Expression ◽  
Stem Cells ◽  
Transcriptional Regulation ◽  
Mrna Stability ◽  
Regulation Of Gene Expression ◽  
Embryonic Stem ◽  
Histone Lysine ◽  
Histone Lysine Methyltransferase ◽  
Lysine Methyltransferase ◽  
Post Transcriptional Regulation

Abstract Embryonic stem cells (ESCs) fate is regulated both at transcriptional and post-transcriptional levels. Indeed, several studies showed that, in addition to gene transcription, mRNA stability and protein synthesis are finely tuned and strongly control the ESCs pluripotency and fate changes. An increasing number of RNA-binding proteins (RBPs) involved in post-transcriptional and translational regulation of gene expression has been identified as regulators of ESC identity. The major lysine methyltransferase Setdb1 is essential for the self-renewal and viability of ESCs. Setdb1 was primarily known to methylate the lysine 9 of histone 3 (H3K9) in the nucleus, where it regulates chromatin functions. However, Setdb1 is also massively localized in the cytoplasm, including in mouse ESCs, where its role remains unknown. Here we show that the cytoplasmic Setdb1 (cSetdb1) is essential for the survival of mESCs. Functional assays further demonstrate that cSetdb1 regulates gene expression post-transcriptionally, affecting the abundance of mRNAs and the rate of newly synthetized proteins. A yeast-two-hybrid assay shows that cSetdb1 interacts with several regulators of mRNA stability and protein translation machinery, such as the ESCs-specific E3 ubiquitin ligase and mRNA silencer Trim71/Lin41. Finally, proteomic analyses reveal that cSetdb1 is required for the integrity of Trim71 complexes involved in mRNA metabolism and translation. Altogether, our data uncover the essential cytoplasmic function of a firstly supposed nuclear “histone” lysine methyltransferase, Setdb1, and provide new insights into the cytoplasmic/post-transcriptional regulation of gene expression mediated by a key epigenetic regulator.

scholarly journals Lin28-mediated post-transcriptional regulation of Oct4 expression in human embryonic stem cells

2009 ◽  
Vol 38 (4) ◽  
pp. 1240-1248 ◽  
Author(s):  
Caihong Qiu ◽  
Yinghong Ma ◽  
Jianquan Wang ◽  
Shuping Peng ◽  
Yingqun Huang
Keyword(s):  
Stem Cells ◽  
Transcriptional Regulation ◽  
Embryonic Stem Cells ◽  
Human Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Oct4 Expression ◽  
Post Transcriptional Regulation

scholarly journals MicroRNA 630 Represses NANOG Expression through Transcriptional and Post-Transcriptional Regulation in Human Embryonal Carcinoma Cells

2021 ◽  
Vol 23 (1) ◽  
pp. 46
Author(s):  
Wing-Keung Chu ◽  
Li-Man Hung ◽  
Chun-Wei Hou ◽  
Jan-Kan Chen
Keyword(s):  
Transcriptional Regulation ◽  
Embryonal Carcinoma ◽  
Carcinoma Cell ◽  
Embryonic Stem ◽  
Embryonal Carcinoma Cell ◽  
Pluripotency Factors ◽  
Nanog Expression ◽  
A Genome ◽  
Target Sites ◽  
Post Transcriptional Regulation

The pluripotent transcription factor NANOG is essential for maintaining embryonic stem cells and driving tumorigenesis. We previously showed that PKC activity is involved in the regulation of NANOG expression. To explore the possible involvement of microRNAs in regulating the expression of key pluripotency factors, we performed a genome-wide analysis of microRNA expression in the embryonal carcinoma cell line NT2/D1 in the presence of the PKC activator, PMA. We found that MIR630 was significantly upregulated in PMA-treated cells. Experimentally, we showed that transfection of MIR630 mimic into embryonal carcinoma cell lines directly targeted the 3′UTR of OCT4, SOX2, and NANOG and markedly suppressed their expression. RNAhybrid and RNA22 algorithms were used to predict miRNA target sites in the NANOG 3’UTR, four possible target sites of MIR630 were identified. To examine the functional interaction between MIR630 and NANOG mRNA, the predicted MIR630 target sites in the NANOG 3’UTR were deleted and the activity of the reporters were compared. After targeted mutation of the predicted MIR630 target sites, the MIR630 mimic inhibited NANOG significantly less than the wild-type reporters. It is worth noting that mutation of a single putative binding site in the 3’UTR of NANOG did not completely abolish MIR630-mediated suppression, suggesting that MIR630 in the NANOG 3’UTR may have multiple binding sites and act together to maximally repress NANOG expression. Interestingly, MIR630 mimics significantly downregulated NANOG gene transcription. Exogenous expression of OCT4, SOX2, and NANOG lacking the 3’UTR almost completely rescued the reduced transcriptional activity of MIR630. MIR630 mediated the expression of differentiation markers in NT2/D1 cells, suggesting that MIR630 leads to the differentiation of NT2/D1 cell. Our findings show that MIR630 represses NANOG through transcriptional and post-transcriptional regulation, suggesting a direct link between core pluripotency factors and MIR630.

scholarly journals Dynamic post-transcriptional regulation during embryonic stem cell differentiation

2017 ◽  
Author(s):  
Patrick R. van den Berg ◽  
Bogdan Budnik ◽  
Nikolai Slavov ◽  
Stefan Semrau
Keyword(s):  
Stem Cells ◽  
Transcriptional Regulation ◽  
Kinetic Model ◽  
Embryonic Stem ◽  
Stem Cell Differentiation ◽  
Specific Gene ◽  
Data Set ◽  
Protein Levels ◽  
Comprehensive Data ◽  
Post Transcriptional Regulation

SummaryDuring in vitro differentiation, pluripotent stem cells undergo extensive remodeling of their gene expression profile. While studied extensively at the transcriptome level, much less is known about protein dynamics. Here, we measured mRNA and protein levels of 7459 genes during differentiation of embryonic stem cells (ESCs). This comprehensive data set revealed pervasive discordance between mRNA and protein. The high temporal resolution of the data made it possible to determine protein turnover rates genome-wide by fitting a kinetic model. This model further enabled us to systematically identify dynamic post-transcriptional regulation. Moreover, we linked different modes of regulation to the function of specific gene sets. Finally, we showed that the kinetic model can be applied to singlecell transcriptomics data to predict protein levels in differentiated cell types. In conclusion, our comprehensive data set, easily accessible through a web application, is a valuable resource for the discovery of post-transcriptional regulation in ESC differentiation.

43-OR: Post-transcriptional Regulation of Slc16a1 mRNA Is Essential for Maintaining Normal ß-Cell Function

Diabetes ◽  
2019 ◽  
Vol 68 (Supplement 1) ◽  
pp. 43-OR
Author(s):  
DINA MOSTAFA ◽  
AKINORI TAKAHASHI ◽  
TADASHI YAMAMOTO
Keyword(s):  
Transcriptional Regulation ◽  
Cell Function ◽  
Post Transcriptional Regulation
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