TY - JOUR
T1 - A nicotinamide phosphoribosyltransferase–GAPDH interaction sustains the stress-induced NMN/NAD+ salvage pathway in the nucleus
AU - Grolla, Ambra A.
AU - Miggiano, Riccardo
AU - Marino, Daniele Di
AU - Bianchi, Michele
AU - Gori, Alessandro
AU - Orsomando, Giuseppe
AU - Gaudino, Federica
AU - Galli, Ubaldina
AU - Grosso, Erika Del
AU - Mazzola, Francesca
AU - Angeletti, Carlo
AU - Guarneri, Martina
AU - Torretta, Simone
AU - Calabrò, Marta
AU - Boumya, Sara
AU - Fan, Xiaorui
AU - Colombo, Giorgia
AU - Travelli, Cristina
AU - Rocchio, Francesca
AU - Aronica, Eleonora
AU - Wohlschlegel, James A.
AU - Deaglio, Silvia
AU - Rizzi, Menico
AU - Genazzani, Armando A.
AU - Garavaglia, Silvia
PY - 2020/3/13
Y1 - 2020/3/13
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=85081966881&partnerID=8YFLogxK
U2 - https://doi.org/10.1074/jbc.RA119.010571
DO - https://doi.org/10.1074/jbc.RA119.010571
M3 - Article
C2 - 31988240
SN - 0021-9258
VL - 295
SP - 3635
EP - 3651
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 11
ER -