A nicotinamide phosphoribosyltransferase–GAPDH interaction sustains the stress-induced NMN/NAD+ salvage pathway in the nucleus

• Published in: The Journal of biological chemistry 2020-03, Vol.295 (11), p.3635-3651
• Main Authors: Grolla, Ambra A., Miggiano, Riccardo, Di Marino, Daniele, Bianchi, Michele, Gori, Alessandro, Orsomando, Giuseppe, Gaudino, Federica, Galli, Ubaldina, Del Grosso, Erika, Mazzola, Francesca, Angeletti, Carlo, Guarneri, Martina, Torretta, Simone, Calabrò, Marta, Boumya, Sara, Fan, Xiaorui, Colombo, Giorgia, Travelli, Cristina, Rocchio, Francesca, Aronica, Eleonora, Wohlschlegel, James A., Deaglio, Silvia, Rizzi, Menico, Genazzani, Armando A., Garavaglia, Silvia
• Format: Article
• Language: English
• Subjects: Animals; Cell Line, Tumor; Cell Nucleus - enzymology; cell stress; GAPDH; Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating) - metabolism; HeLa Cells; Humans; Kinetics; melanoma; Melanoma, Experimental - enzymology; Melanoma, Experimental - pathology; Metabolism; Mice; NAD - metabolism; NAD biosynthesis; NAD compartmentalization; NAMPT; nicotinamide adenine dinucleotide (NAD); nicotinamide mononucleotide (NMN); Nicotinamide Mononucleotide - chemistry; Nicotinamide Mononucleotide - metabolism; Nicotinamide Phosphoribosyltransferase - chemistry; Nicotinamide Phosphoribosyltransferase - metabolism; NIH 3T3 Cells; NMN/NAD+ salvage pathway; nucleus; Protein Binding; Protein Multimerization; Protein Transport; protein-protein interaction; redox cycling; Stress, Physiological
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.RA119.010571

All cells require sustained intracellular energy flux, which is driven by redox chemistry at the subcellular level. NAD+, its phosphorylated variant NAD(P)+, and its reduced forms NAD(P)/NAD(P)H are all redox cofactors with key roles in energy metabolism and are substrates for several NAD-consuming enzymes (e.g. poly(ADP-ribose) polymerases, sirtuins, and others). The nicotinamide salvage pathway, constituted by nicotinamide mononucleotide adenylyltransferase (NMNAT) and nicotinamide phosphoribosyltransferase (NAMPT), mainly replenishes NAD+ in eukaryotes. However, unlike NMNAT1, NAMPT is not known to be a nuclear protein, prompting the question of how the nuclear NAD+ pool is maintained and how it is replenished upon NAD+ consumption. In the present work, using human and murine cells; immunoprecipitation, pulldown, and surface plasmon resonance assays; and immunofluorescence, small-angle X-ray scattering, and MS-based analyses, we report that GAPDH and NAMPT form a stable complex that is essential for nuclear translocation of NAMPT. This translocation furnishes NMN to replenish NAD+ to compensate for the activation of NAD-consuming enzymes by stressful stimuli induced by exposure to H2O2 or S-nitrosoglutathione and DNA damage inducers. These results indicate that by forming a complex with GAPDH, NAMPT can translocate to the nucleus and thereby sustain the stress-induced NMN/NAD+ salvage pathway.