Autoradiographic studies of nicotinic acid utilization in human-mouse heterokaryons and inhibition of utilization in newly-formed hybrid cells.
Although most mammalian cell lines can utilize either nicotinic acid or nicotinamide for the biosynthesis of nicotinamide adenine dinucleotide (NAD), thymidine kinase-deficient, mouse 3T3-4F cells are unable to utilize nicotinic acid. When 3T3-4E cells were fused with human D98/AH2 cells, autoradiography showed that the resultant heterokaryons synthesized NAD from nicotinic acid at rates comparable to the human parental cell. The rate of nicotinic acid utilization in heterokaryons remained unchanged over the four-day period of study following cell fusion. In contrast to the results observed with heterokaryons, nicotinic acid utilization was markedly reduced in hybrid cells. Of 100 hybrid clones examined at four or five days following cell fusion, 60 utilized nicotinic acid at rates less than one tenth that of the parental human cell. Similar results were observed in hybrid clones at nine or ten days following fusion. Uniformly high rates of NAD biosynthesis were observed in hybrid clones with nicotinamide as the precursor. This excludes the possibility that the reduction in nicotinic acid utilization in hybrid cells is due to a general metabolic dysfunction. The biochemical mechanism by which nicotinic acid utilization is markedly reduced has not been determined with certainty, however, several observations suggest genetic suppression.
D000630:Aminopterin / Q000494:pharmacology; D002455:Cell Division / Q000187:drug effects; D002459:Cell Fusion; D003470:Culture Media; D004247:DNA / Q000096:biosynthesis; D006822:Hybrid Cells / Q000378:metabolism*; D009243:NAD / Q000096:biosynthesis; D009536:Niacinamide / Q000378:metabolism; D009539:Nicotinic Acids / Q000378:metabolism*
D003470:Culture Media; D009539:Nicotinic Acids; D009243:NAD; D009536:Niacinamide; D004247:DNA; D000630:Aminopterin