Lysosomal Signaling Licenses Embryonic Stem Cell Differentiation via Inactivation of Tfe3

Florian Villegas, Daphné Lehalle, Daniela Mayer, Melanie Rittirsch, Michael B. Stadler, Marietta Zinner, Daniel Olivieri, Pierre Vabres, Laurence Duplomb-Jego, Eveline S. J. M. de Bont, Yannis Duffourd, Floor Duijkers, Magali Avila, David Geneviève, Nada Houcinat, Thibaud Jouan, Paul Kuentz, Klaske D. Lichtenbelt, Christel Thauvin-Robinet, Judith St-OngeJulien Thevenon, Koen L. I. van Gassen, Mieke van Haelst, Silvana van Koningsbruggen, Daniel Hess, Sebastien A. Smallwood, Jean-Baptiste Rivière, Laurence Faivre, Joerg Betschinger

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Self-renewal and differentiation of pluripotent murine embryonic stem cells (ESCs) is regulated by extrinsic signaling pathways. It is less clear whether cellular metabolism instructs developmental progression. In an unbiased genome-wide CRISPR/Cas9 screen, we identified components of a conserved amino-acid-sensing pathway as critical drivers of ESC differentiation. Functional analysis revealed that lysosome activity, the Ragulator protein complex, and the tumor-suppressor protein Folliculin enable the Rag GTPases C and D to bind and seclude the bHLH transcription factor Tfe3 in the cytoplasm. In contrast, ectopic nuclear Tfe3 represses specific developmental and metabolic transcriptional programs that are associated with peri-implantation development. We show differentiation-specific and non-canonical regulation of Rag GTPase in ESCs and, importantly, identify point mutations in a Tfe3 domain required for cytoplasmic inactivation as potentially causal for a human developmental disorder. Our work reveals an instructive and biomedically relevant role of metabolic signaling in licensing embryonic cell fate transitions.
Original languageEnglish
Pages (from-to)257-270.e8
JournalCell Stem Cell
Issue number2
Publication statusPublished - 7 Feb 2019
Externally publishedYes

Cite this

Villegas, F., Lehalle, D., Mayer, D., Rittirsch, M., Stadler, M. B., Zinner, M., ... Betschinger, J. (2019). Lysosomal Signaling Licenses Embryonic Stem Cell Differentiation via Inactivation of Tfe3. Cell Stem Cell, 24(2), 257-270.e8.