TY - JOUR
T1 - Exit from pluripotency is gated by intracellular redistribution of the bHLH transcription factor Tfe3
AU - Betschinger, Joerg
AU - Nichols, Jennifer
AU - Dietmann, Sabine
AU - Corrin, Philip D.
AU - Paddison, Patrick J.
AU - Smith, Austin
N1 - Funding Information:
We thank Tüzer Kalkan and Graziano Martello for help, advice, and reagents; Raja Kittappa, Martin Leeb, and Harry Leitch for comments; Ravi Nookala for discussion; Ge Guo for providing OEC-2 EpiSCs; and Sandra Gómez-López and Jose Silva for sharing CreERT2, pDONR221-Klf4, and -Nanog plasmids. We are grateful to Laura Schmidt for generously providing Flcn antibodies and mice; Kathryn Lilley, Mike Deery, and Svenja Hesters of the Cambridge Centre for Proteomics for performing and analyzing mass spectrometry; Rachael Walker for flow cytometry; William Mansfield and Charles-Etienne Dumeau for blastocyst injections; Samuel Jameson and team for husbandry; Peter Humphreys for image quantification; Maike Paramor for ChIP-Seq library generation; and Rosalind Drummond and Melanie Rittirsch for technical assistance. This study was funded by the Medical Research Council, Wellcome Trust, and the European Commission FP7 project SyBOSS (242129). J.B. is recipient of a Human Frontier Science Program fellowship (LT00686/2007-L), P.J.P. is a Pew Scholar in Biomedical Sciences, and A.S. is a Medical Research Council Professor. J.B. performed and interpreted experiments, J.N. performed embryo immunohistochemistry and ESC derivations, S.D. performed bioinformatical analysis, and P.D.C. and P.J.P. provided and assisted with the siRNA-screening platform. A.S. supervised the study and wrote the paper with J.B.
PY - 2013/4/11
Y1 - 2013/4/11
N2 - Factors that sustain self-renewal of mouse embryonic stem cells (ESCs) are well described. In contrast, the machinery regulating exit from pluripotency is ill defined. In a large-scale small interfering RNA (siRNA) screen, we found that knockdown of the tumor suppressors Folliculin (Flcn) and Tsc2 prevent ESC commitment. Tsc2 lies upstream of mammalian target of rapamycin (mTOR), whereas Flcn acts downstream and in parallel. Flcn with its interaction partners Fnip1 and Fnip2 drives differentiation by restricting nuclear localization and activity of the bHLH transcription factor Tfe3. Conversely, enforced nuclear Tfe3 enables ESCs to withstand differentiation conditions. Genome-wide location and functional analyses showed that Tfe3 directly integrates into the pluripotency circuitry through transcriptional regulation of Esrrb. These findings identify a cell-intrinsic rheostat for destabilizing ground-state pluripotency to allow lineage commitment. Congruently, stage-specific subcellular relocalization of Tfe3 suggests that Flcn-Fnip1/2 contributes to developmental progression of the pluripotent epiblast in vivo.
AB - Factors that sustain self-renewal of mouse embryonic stem cells (ESCs) are well described. In contrast, the machinery regulating exit from pluripotency is ill defined. In a large-scale small interfering RNA (siRNA) screen, we found that knockdown of the tumor suppressors Folliculin (Flcn) and Tsc2 prevent ESC commitment. Tsc2 lies upstream of mammalian target of rapamycin (mTOR), whereas Flcn acts downstream and in parallel. Flcn with its interaction partners Fnip1 and Fnip2 drives differentiation by restricting nuclear localization and activity of the bHLH transcription factor Tfe3. Conversely, enforced nuclear Tfe3 enables ESCs to withstand differentiation conditions. Genome-wide location and functional analyses showed that Tfe3 directly integrates into the pluripotency circuitry through transcriptional regulation of Esrrb. These findings identify a cell-intrinsic rheostat for destabilizing ground-state pluripotency to allow lineage commitment. Congruently, stage-specific subcellular relocalization of Tfe3 suggests that Flcn-Fnip1/2 contributes to developmental progression of the pluripotent epiblast in vivo.
UR - http://www.scopus.com/inward/record.url?scp=84876261517&partnerID=8YFLogxK
U2 - 10.1016/j.cell.2013.03.012
DO - 10.1016/j.cell.2013.03.012
M3 - Article
C2 - 23582324
AN - SCOPUS:84876261517
SN - 0092-8674
VL - 153
SP - 335
EP - 347
JO - Cell
JF - Cell
IS - 2
ER -