Ammonia transporter RhBG initiates downstream signaling and functional responses by activating NFκB

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 121; no. 31; p. e2314760121
• Main Authors: Mishra, Saurabh, Welch, Nicole, Singh, Shashi Shekhar, Singh, Khuraijam Dhanachandra, Bellar, Annette, Kumar, Avinash, Deutz, Lars N, Hanlon, Maxmillian D, Kant, Sashi, Dastidar, Sumitava, Patel, Hailee, Agrawal, Vandana, Attaway, Amy H, Musich, Ryan, Stark, George R, Tedesco, Francesco Saverio, Truskey, George A, Weiner, I David, Karnik, Sadashiva S, Dasarathy, Srinivasan
• Format: Journal Article
• Language: English
• Published: United States National Academy of Sciences 30.07.2024
• Subjects: Ammonia; Ammonia - metabolism; Animals; Blood groups; Cytotoxicity; Decoupling; Glycoproteins; Hepatocytes; Human influences; Humans; Hyperammonemia; Hyperammonemia - genetics; Hyperammonemia - metabolism; Interleukin 1; Intracellular; Intracellular signalling; Mammals; Membrane Transport Proteins - genetics; Membrane Transport Proteins - metabolism; Metabolites; Mice; Mice, Knockout; Muscle, Skeletal - metabolism; Muscles; Musculoskeletal system; MyD88 protein; Myeloid Differentiation Factor 88 - genetics; Myeloid Differentiation Factor 88 - metabolism; Myotubes; NF-kappa B - metabolism; NF-κB protein; Pluripotency; Signal Transduction; Skeletal muscle; Stem cells; Substrates; Toll-like receptors; Transmembrane domains; Urea; hyperammonemia; NFkB; solute transporter; transceptor; skeletal muscle
• ISSN: 0027-8424; 1091-6490; 1091-6490
• DOI: 10.1073/pnas.2314760121

Transceptors, solute transporters that facilitate intracellular entry of molecules and also initiate intracellular signaling events, have been primarily studied in lower-order species. Ammonia, a cytotoxic endogenous metabolite, is converted to urea in hepatocytes for urinary excretion in mammals. During hyperammonemia, when hepatic metabolism is impaired, nonureagenic ammonia disposal occurs primarily in skeletal muscle. Increased ammonia uptake in skeletal muscle is mediated by a membrane-bound, 12 transmembrane domain solute transporter, Rhesus blood group-associated B glycoprotein (RhBG). We show that in addition to its transport function, RhBG interacts with myeloid differentiation primary response-88 (MyD88) to initiate an intracellular signaling cascade that culminates in activation of NFκB. We also show that ammonia-induced MyD88 signaling is independent of the canonical toll-like receptor-initiated mechanism of MyD88-dependent NFκB activation. In silico, in vitro, and in situ experiments show that the conserved cytosolic J-domain of the RhBG protein interacts with the Toll-interleukin-1 receptor (TIR) domain of MyD88. In skeletal muscle from human patients, human-induced pluripotent stem cell-derived myotubes, and myobundles show an interaction of RhBG-MyD88 during hyperammonemia. Using complementary experimental and multiomics analyses in murine myotubes and mice with muscle-specific RhBG or MyD88 deletion, we show that the RhBG-MyD88 interaction is essential for the activation of NFkB but not ammonia transport. Our studies show a paradigm of substrate-dependent regulation of transceptor function with the potential for modulation of cellular responses in mammalian systems by decoupling transport and signaling functions of transceptors.