Premium
Reactivation of NAD(H) biosynthetic pathway by exogenous NAD + in nil cells severely depleted of NAD(H)
Author(s) -
Mandel Kenneth G.,
Lively Mary K.,
Lombardi Donald,
Amos Harold
Publication year - 1983
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/jcp.1041140214
Subject(s) - nad+ kinase , intracellular , nicotinamide , nicotinamide mononucleotide , nicotinamide adenine dinucleotide , biochemistry , biology , niacinamide , chemistry , microbiology and biotechnology , enzyme
The culture of Nil hamster fibroblasts in MEM lacking nicotinamide (NAm − MEM) leads to: (1) the rapid loss of intracellular total nicotinamide adenine dinucleotide (NAD(H)) content in these cells from a level of 150–200 pmoles/10 5 cells to less than 20 pmoles/10 5 cells; (2) the cessation of cell division and inhibition of DNA synthesis; and (3) a reduction of glucose consumption and lactic acid production. In most situations, following nicotinamide starvation, the restoration of intracellular NAD(H) follows rapidly the readdition of NAD + (oxidized), nicotinamide mononucleotide (NMN), nicotinamide, or nicotinic acid. Resumption of cell division occurs after only a lag of about 24 hours. Nil cells subcultured for three consecutive times in the absence of nicotinamide (3° NAm‐ cells) exhibit different behavior. These severely starved cells are incapable of quickly restoring their intracellular NAD(H) content to normal levels when provided with any pyridine ring compound except NAD + . One‐hour exposure of such cells to NAD + allows utilization of nicotinamide to rapidly restore intracellular NAD(H). This short incubation with NAD + does not result in any significant restoration of intracellular NAD(H) or lead to the accumulation of an intracellular pool of some precursor. This function of NAD + as a stimulatory signal to the NAD(H)‐biosynthetic pathway in severely starved Nil cells is a previously unreported role of NAD + , and does not require protein synthesis.