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

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