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Development of a mouse model expressing a bifunctional glutathione-synthesizing enzyme to study glutathione limitation in vivo.
Timson, Rebecca C; Khan, Artem; Uygur, Beste; Saad, Marwa; Yeh, Hsi-Wen; DelGaudio, Nicole L; Weber, Ross; Alwaseem, Hanan; Gao, Jing; Yang, Chingwen; Birsoy, Kivanç.
Afiliación
  • Timson RC; Laboratory of Metabolic Regulation and Genetics, The Rockefeller University, New York, New York, USA.
  • Khan A; Laboratory of Metabolic Regulation and Genetics, The Rockefeller University, New York, New York, USA.
  • Uygur B; Laboratory of Metabolic Regulation and Genetics, The Rockefeller University, New York, New York, USA.
  • Saad M; Laboratory of Mucosal Immunology, The Rockefeller University, New York, New York, USA.
  • Yeh HW; Laboratory of Metabolic Regulation and Genetics, The Rockefeller University, New York, New York, USA.
  • DelGaudio NL; Laboratory of Metabolic Regulation and Genetics, The Rockefeller University, New York, New York, USA.
  • Weber R; Department of Radiation Oncology, Memorial Sloan Kettering Cancer Center, New York, New York, USA.
  • Alwaseem H; The Proteomics Resource Center, The Rockefeller University, New York, New York, USA.
  • Gao J; The CRISPR & Genome Editing Center, The Rockefeller University, New York, New York, USA.
  • Yang C; The CRISPR & Genome Editing Center, The Rockefeller University, New York, New York, USA.
  • Birsoy K; Laboratory of Metabolic Regulation and Genetics, The Rockefeller University, New York, New York, USA. Electronic address: kbirsoy@rockefeller.edu.
J Biol Chem ; 300(2): 105645, 2024 Feb.
Article en En | MEDLINE | ID: mdl-38218225
ABSTRACT
Glutathione (GSH) is a highly abundant tripeptide thiol that performs diverse protective and biosynthetic functions in cells. While changes in GSH availability are associated with inborn errors of metabolism, cancer, and neurodegenerative disorders, studying the limiting role of GSH in physiology and disease has been challenging due to its tight regulation. To address this, we generated cell and mouse models that express a bifunctional glutathione-synthesizing enzyme from Streptococcus thermophilus (GshF), which possesses both glutamate-cysteine ligase and glutathione synthase activities. GshF expression allows efficient production of GSH in the cytosol and mitochondria and prevents cell death in response to GSH depletion, but not ferroptosis induction, indicating that GSH is not a limiting factor under lipid peroxidation. CRISPR screens using engineered enzymes further revealed genes required for cell proliferation under cellular and mitochondrial GSH depletion. Among these, we identified the glutamate-cysteine ligase modifier subunit, GCLM, as a requirement for cellular sensitivity to buthionine sulfoximine, a glutathione synthesis inhibitor. Finally, GshF expression in mice is embryonically lethal but sustains postnatal viability when restricted to adulthood. Overall, our work identifies a conditional mouse model to investigate the limiting role of GSH in physiology and disease.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Glutamato-Cisteína Ligasa / Glutatión Tipo de estudio: Prognostic_studies Límite: Animals / Humans Idioma: En Revista: J Biol Chem Año: 2024 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Estados Unidos
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Glutamato-Cisteína Ligasa / Glutatión Tipo de estudio: Prognostic_studies Límite: Animals / Humans Idioma: En Revista: J Biol Chem Año: 2024 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Estados Unidos