Regulation of Hematopoiesis and Methionine Homeostasis by mTORC1 Inhibitor NPRL2

• Published in: Cell reports (Cambridge) Vol. 12; no. 3; pp. 371 - 379
• Main Authors: Dutchak, Paul A., Laxman, Sunil, Estill, Sandi Jo, Wang, Chensu, Wang, Yun, Wang, Yiguang, Bulut, Gamze B., Gao, Jinming, Huang, Lily J., Tu, Benjamin P.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 21-07-2015; Elsevier
• Subjects: Animals; Female; Hematopoiesis - physiology; Mechanistic Target of Rapamycin Complex 1; Methionine - metabolism; Mice; Mice, Inbred C57BL; Mice, Knockout; Multiprotein Complexes - antagonists & inhibitors; Multiprotein Complexes - metabolism; TOR Serine-Threonine Kinases - antagonists & inhibitors; TOR Serine-Threonine Kinases - metabolism; Tumor Suppressor Proteins - metabolism
• ISSN: 2211-1247; 2211-1247
• DOI: 10.1016/j.celrep.2015.06.042

Nitrogen permease regulator-like 2 (NPRL2) is a component of a conserved complex that inhibits mTORC1 (mammalian Target Of Rapamycin Complex 1) in response to amino acid insufficiency. Here, we show that NPRL2 is required for mouse viability and that its absence significantly compromises fetal liver hematopoiesis in developing embryos. Moreover, NPRL2 KO embryos have significantly reduced methionine levels and exhibit phenotypes reminiscent of cobalamin (vitamin B12) deficiency. Consistent with this idea, NPRL2 KO liver and mouse embryonic fibroblasts (MEFs) show defective processing of the cobalamin-transport protein transcobalamin 2, along with impaired lysosomal acidification and lysosomal gene expression. NPRL2 KO MEFs exhibit a significant defect in the cobalamin-dependent synthesis of methionine from homocysteine, which can be rescued by supplementation with cyanocobalamin. Taken together, these findings demonstrate a role for NPRL2 and mTORC1 in the regulation of lysosomal-dependent cobalamin processing, methionine synthesis, and maintenance of cellular re-methylation potential, which are important during hematopoiesis.
[Display omitted]
•Loss of NPRL2 results in defective liver hematopoiesis in the developing embryo•NPRL2 is necessary to repress mTORC1 activity during nutrient limitation•NPRL2 KO compromises lysosomal acidification•Lysosomal processing of cobalamin and methionine synthase are defective in NPRL2 KO
NPRL2 is a constituent of the conserved GATOR1 complex that inhibits mTORC1 signaling during amino acid starvation. Dutchak et al. show that loss of NPRL2 results in defective fetal liver hematopoiesis and compromises lysosomal acidification and processing of cobalamin, an essential cofactor for methionine synthase.