Publish in OALib Journal
ISSN: 2333-9721
APC: Only $99

Paper Submission

Views	Downloads

Relative Articles
Transcriptional analysis of early lineage commitment in human embryonic stem cells
Transcriptional Regulation of Lineage Commitment - A Stochastic Model of Cell Fate Decisions
Cell Hierarchy and Lineage Commitment in the Bovine Mammary Gland
Notch Promotes Neural Lineage Entry by Pluripotent Embryonic Stem Cells
Notch Promotes Neural Lineage Entry by Pluripotent Embryonic Stem Cells
Activation of GSK3β by Sirt2 Is Required for Early Lineage Commitment of Mouse Embryonic Stem Cell
Integrating Extrinsic and Intrinsic Cues into a Minimal Model of Lineage Commitment for Hematopoietic Progenitors
A Continuum of Cell States Spans Pluripotency and Lineage Commitment in Human Embryonic Stem Cells
Feedback Signals in Myelodysplastic Syndromes: Increased Self-Renewal of the Malignant Clone Suppresses Normal Hematopoiesis
Microenvironment Modulates Osteogenic Cell Lineage Commitment in Differentiated Embryonic Stem Cells
More...

PLOS ONE 2013

ERK2 Suppresses Self-Renewal Capacity of Embryonic Stem Cells, but Is Not Required for Multi-Lineage Commitment

DOI: 10.1371/journal.pone.0060907

William B. Hamilton, Keisuke Kaji, Tilo Kunath

Full-Text Cite this paper Add to My Lib

Abstract:

Activation of the FGF-ERK pathway is necessary for na？ve mouse embryonic stem (ES) cells to exit self-renewal and commit to early differentiated lineages. Here we show that genetic ablation of Erk2, the predominant ERK isozyme expressed in ES cells, results in hyper-phosphorylation of ERK1, but an overall decrease in total ERK activity as judged by substrate phosphorylation and immediate-early gene (IEG) induction. Normal induction of this subset of canonical ERK targets, as well as p90RSK phosphorylation, was rescued by transgenic expression of either ERK1 or ERK2 indicating a degree of functional redundancy. In contrast to previously published work, Erk2-null ES cells exhibited no detectable defect in lineage specification to any of the three germ layers when induced to differentiate in either embryoid bodies or in defined neural induction conditions. However, under self-renewing conditions Erk2-null ES cells express increased levels of the pluripotency-associated transcripts, Nanog and Tbx3, a decrease in Nanog-GFP heterogeneity, and exhibit enhanced self-renewal in colony forming assays. Transgenic add-back of ERK2 is capable of restoring normal pluripotent gene expression and self-renewal capacity. We show that ERK2 contributes to the destabilization of ES cell self-renewal by reducing expression of pluripotency genes, such as Nanog, but is not specifically required for the early stages of germ layer specification.

References

[1]	Brook FA, Gardner RL (1997) The origin and efficient derivation of embryonic stem cells in the mouse. Proc Natl Acad Sci USA 94: 5709–5712.
[2]	Evans MJ, Kaufman MH (1981) Establishment in culture of pluripotential cells from mouse embryos. Nature 292: 154–156.
[3]	Martin GR (1981) Isolation of a pluripotent cell line from early mouse embryos cultured in medium conditioned by teratocarcinoma stem cells. Proc Natl Acad Sci USA 78: 7634–7638.
[4]	Robertson E, Bradley A, Kuehn M, Evans M (1986) Germ-line transmission of genes introduced into cultured pluripotential cells by retroviral vector. Nature 323: 445–448 doi:10.1038/323445a0.
[5]	Ying Q-L, Wray J, Nichols J, Batlle-Morera L, Doble B, et al. (2008) The ground state of embryonic stem cell self-renewal. Nature 453: 519–523 doi:10.1038/nature06968.
[6]	Najm FJ, Chenoweth JG, Anderson PD, Nadeau JH, Redline RW, et al. (2011) Isolation of epiblast stem cells from preimplantation mouse embryos. Cell Stem Cell 8: 318–325 doi:10.1016/j.stem.2011.01.016.
[7]	Ying Q-L, Nichols J, Chambers I, Smith A (2003) BMP induction of Id proteins suppresses differentiation and sustains embryonic stem cell self-renewal in collaboration with STAT3. Cell 115: 281–292.
[8]	Avilion AAA, Nicolis SKS, Pevny LHL, Perez LL, Vivian NN, et al. (2003) Multipotent cell lineages in early mouse development depend on SOX2 function. Genes Dev 17: 126–140 doi:10.1101/gad.224503.
[9]	Niwa H, Miyazaki J, Smith AG (2000) Quantitative expression of Oct-3/4 defines differentiation, dedifferentiation or self-renewal of ES cells. Nat Genet 24: 372–376 doi:10.1038/74199.
[10]	Loh Y-H, Wu Q, Chew J-L, Vega VB, Zhang W, et al. (2006) The Oct4 and Nanog transcription network regulates pluripotency in mouse embryonic stem cells. Nat Genet 38: 431–440 doi:10.1038/ng1760.
[11]	Niwa H, Ogawa K, Shimosato D, Adachi K (2009) A parallel circuit of LIF signalling pathways maintains pluripotency of mouse ES cells. Nature 460: 118–122 doi:10.1038/nature08113.
[12]	Silva J, Smith A (2008) Capturing pluripotency. Cell 132: 532–536 doi:10.1016/j.cell.2008.02.006.
[13]	Arman E, Haffner-Krausz R, Chen Y, Heath JK, Lonai P (1998) Targeted disruption of fibroblast growth factor (FGF) receptor 2 suggests a role for FGF signaling in pregastrulation mammalian development. Proc Natl Acad Sci USA 95: 5082–5087.
[14]	Chazaud C, Yamanaka Y, Pawson T, Rossant J (2006) Early lineage segregation between epiblast and primitive endoderm in mouse blastocysts through the Grb2-MAPK pathway. Developmental Cell 10: 615–624 doi:10.1016/j.devcel.2006.02.020.
[15]	Cheng AM, Saxton TM, Sakai R, Kulkarni S, Mbamalu G, et al. (1998) Mammalian Grb2 regulates multiple steps in embryonic development and malignant transformation. Cell 95: 793–803.
[16]	Feldman B, Poueymirou W, Papaioannou VE, DeChiara TM, Goldfarb M (1995) Requirement of FGF-4 for postimplantation mouse development. Science 267: 246–249.
[17]	Yuan H, Corbi N, Basilico C, Dailey L (1995) Developmental-specific activity of the FGF-4 enhancer requires the synergistic action of Sox2 and Oct-3. Genes Dev 9: 2635–2645.
[18]	Kunath T, Saba-El-Leil MK, Almousailleakh M, Wray J, Meloche S, et al. (2007) FGF stimulation of the Erk1/2 signalling cascade triggers transition of pluripotent embryonic stem cells from self-renewal to lineage commitment. Development 134: 2895–2902 doi:10.1242/dev.02880.
[19]	Wilder PJ, Kelly D, Brigman K, Peterson CL, Nowling T, et al. (1997) Inactivation of the FGF-4 gene in embryonic stem cells alters the growth and/or the survival of their early differentiated progeny. Dev Biol 192: 614–629 doi:10.1006/dbio.1997.8777.
[20]	Nichols J, Jones K, Phillips JM, Newland SA, Roode M, et al. (2009) Validated germline-competent embryonic stem cell lines from nonobese diabetic mice. Nat Med 15: 814–818 doi:10.1038/nm.1996.
[21]	Buehr M, Meek S, Blair K, Yang J, Ure J, et al. (2008) Capture of authentic embryonic stem cells from rat blastocysts. Cell 135: 1287–1298 doi:10.1016/j.cell.2008.12.007.
[22]	Li P, Tong C, Mehrian-Shai R, Jia L, Wu N, et al. (2008) Germline competent embryonic stem cells derived from rat blastocysts. Cell 135: 1299–1310 doi:10.1016/j.cell.2008.12.006.
[23]	Saba-El-Leil MK, Vella FDJ, Vernay B, Voisin L, Chen L, et al. (2003) An essential function of the mitogen-activated protein kinase Erk2 in mouse trophoblast development. EMBO Rep 4: 964–968 doi:10.1038/sj.embor.embor939.
[24]	Yao Y, Li W, Wu J, Germann UA, Su MSS, et al. (2003) Extracellular signal-regulated kinase 2 is necessary for mesoderm differentiation. Proc Natl Acad Sci USA 100: 12759–12764 doi:10.1073/pnas.2134254100.
[25]	Fischer AM, Katayama CD, Pagès G, Pouysségur J, Hedrick SM (2005) The role of erk1 and erk2 in multiple stages of T cell development. Immunity 23: 431–443 doi:10.1016/j.immuni.2005.08.013.
[26]	Dalby KN, Morrice N, Caudwell FB, Avruch J, Cohen P (1998) Identification of regulatory phosphorylation sites in mitogen-activated protein kinase (MAPK)-activated protein kinase-1a/p90rsk that are inducible by MAPK. J Biol Chem 273: 1496–1505.
[27]	Santos SDM, Verveer PJ, Bastiaens PIH (2007) Growth factor-induced MAPK network topology shapes Erk response determining PC-12 cell fate. Nat Cell Biol 9: 324–330 doi:10.1038/ncb1543.
[28]	De Cesare D, Jacquot S, Hanauer A, Sassone-Corsi P (1998) Rsk-2 activity is necessary for epidermal growth factor-induced phosphorylation of CREB protein and transcription of c-fos gene. Proc Natl Acad Sci USA 95: 12202–12207.
[29]	Nakakuki T, Birtwistle MR, Saeki Y, Yumoto N, Ide K, et al. (2010) Ligand-specific c-Fos expression emerges from the spatiotemporal control of ErbB network dynamics. Cell 141: 884–896 doi:10.1016/j.cell.2010.03.054.
[30]	Choudhury GG (2001) Akt serine threonine kinase regulates platelet-derived growth factor-induced DNA synthesis in glomerular mesangial cells: regulation of c-fos AND p27(kip1) gene expression. J Biol Chem 276: 35636–35643 doi:10.1074/jbc.M100946200.
[31]	Brondello JM, Pouysségur J, McKenzie FR (1999) Reduced MAP kinase phosphatase-1 degradation after p42/p44MAPK-dependent phosphorylation. Science 286: 2514–2517.
[32]	Brondello JM, Brunet A, Pouysségur J, McKenzie FR (1997) The dual specificity mitogen-activated protein kinase phosphatase-1 and -2 are induced by the p42/p44MAPK cascade. J Biol Chem 272: 1368–1376.
[33]	Murphy LO, Smith S, Chen R-H, Fingar DC, Blenis J (2002) Molecular interpretation of ERK signal duration by immediate early gene products. Nat Cell Biol 4: 556–564 doi:10.1038/ncb822.
[34]	White J, Dalton S (2005) Cell cycle control of embryonic stem cells. Stem Cell Rev 1: 131–138 doi:10.1385/SCR:1:2:131.
[35]	Ying Q-L, Stavridis M, Griffiths D, Li M, Smith A (2003) Conversion of embryonic stem cells into neuroectodermal precursors in adherent monoculture. 4 doi:10.1038/nbt780.
[36]	Chambers I, Silva J, Colby D, Nichols J, Nijmeijer B, et al. (2007) Nanog safeguards pluripotency and mediates germline development. Nature 450: 1230–1234 doi:10.1038/nature06403.
[37]	Singh AM, Hamazaki T, Hankowski KE, Terada N (2007) A heterogeneous expression pattern for Nanog in embryonic stem cells. Stem Cells 25: 2534–2542 doi:10.1634/stemcells.2007-0126.
[38]	Ivanova N, Dobrin R, Lu R, Kotenko I, Levorse J, et al. (2006) Dissecting self-renewal in stem cells with RNA interference. Nature 442: 533–538 doi:10.1038/nature04915.
[39]	van den Berg DLC, Snoek T, Mullin NP, Yates A, Bezstarosti K, et al. (2010) An Oct4-centered protein interaction network in embryonic stem cells. Cell Stem Cell 6: 369–381 doi:10.1016/j.stem.2010.02.014.
[40]	Chambers I, Colby D, Robertson M, Nichols J, Lee S, et al. (2003) Functional expression cloning of Nanog, a pluripotency sustaining factor in embryonic stem cells. Cell 113: 643–655.
[41]	Kholodenko BN, Hancock JF, Kolch W (2010) Signalling ballet in space and time. Nat Rev Mol Cell Biol 11: 414–426 doi:10.1038/nrm2901.
[42]	Voisin L, Saba-El-Leil MK, Julien C, Frémin C, Meloche S (2010) Genetic demonstration of a redundant role of extracellular signal-regulated kinase 1 (ERK1) and ERK2 mitogen-activated protein kinases in promoting fibroblast proliferation. Mol Cell Biol 30: 2918–2932 doi:10.1128/MCB.00131-10.
[43]	Canham MA, Sharov AA, Ko MSH, Brickman JM (2010) Functional heterogeneity of embryonic stem cells revealed through translational amplification of an early endodermal transcript. PLoS Biol 8: e1000379 doi:10.1371/journal.pbio.1000379.
[44]	Davies OR, Lin C-Y, Radzisheuskaya A, Zhou X, Taube J, et al. (2013) Tcf15 primes pluripotent cells for differentiation. Cell Rep 3: 472–484 doi:10.1016/j.celrep.2013.01.017.
[45]	Burdon T, Stracey C, Chambers I, Nichols J, Smith A (1999) Suppression of SHP-2 and ERK signalling promotes self-renewal of mouse embryonic stem cells. Dev Biol 210: 30–43 doi:10.1006/dbio.1999.9265.
[46]	Niwa H, Yamamura K, Miyazaki J (1991) Efficient selection for high-expression transfectants with a novel eukaryotic vector. Gene 108: 193–199.
[47]	Tsakiridis A, Tzouanacou E, Rahman A, Colby D, Axton R, et al. (2009) Expression-independent gene trap vectors for random and targeted mutagenesis in embryonic stem cells. Nucleic Acids Res 37: e129 doi:10.1093/nar/gkp640.
[48]	Rogers S, Wells R, Rechsteiner M (1986) Amino acid sequences common to rapidly degraded proteins: the PEST hypothesis. Science 234: 364–368.
[49]	Gurtu V, Yan G, Zhang G (1996) IRES bicistronic expression vectors for efficient creation of stable mammalian cell lines. Biochemical and Biophysical Research Communications 229: 295–298 doi:10.1006/bbrc.1996.1795.
[50]	Hooper M, Hardy K, Handyside A, Hunter S, Monk M (1987) HPRT-deficient (Lesch-Nyhan) mouse embryos derived from germline colonization by cultured cells. Nature 326: 292–295 doi:10.1038/326292a0.
[51]	Kiyatkin A, Aksamitiene E (2009) Multistrip western blotting to increase quantitative data output. Methods Mol Biol 536: 149–161 doi:10.1007/978-1-59745-542-8-17.

Full-Text