The inhibition of nucleic acid synthesis by mycophenolic acid

1. Mycophenolic acid, an antibiotic of some antiquity that more recently has been found to have marked activity against a range of tumours in mice and rats, strongly inhibits DNA synthesis in the L strain of fibroblasts in vitro. 2. The extent of the inhibition of DNA synthesis is markedly increased by preincubation of the cells with mycophenolic acid before the addition of [(14)C]thymidine. 3. The inhibition of DNA synthesis by mycophenolic acid in L cells in vitro is reversed by guanine in a non-competitive manner, but not by hypoxanthine, xanthine or adenine. 4. The reversal of inhibition by guanine can be suppressed by hypoxanthine, 6-mercaptopurine and adenine. 5. Mycophenolic acid does not inhibit the incorporation of [(14)C]thymidine into DNA in suspensions of Landschütz and Yoshida ascites cells in vitro. 6. Mycophenolic acid inhibits the conversion of [(14)C]hypoxanthine into cold-acid-soluble and -insoluble guanine nucleotides in Landschütz and Yoshida ascites cells and also in L cells in vitro. There is some increase in the radioactivity of the adenine fraction in the presence of the antibiotic. 7. Mycophenolic acid inhibits the conversion of [(14)C]hypoxanthine into xanthine and guanine fractions in a cell-free system from Landschütz cells capable of converting hypoxanthine into IMP, XMP and GMP. 8. Preparations of IMP dehydrogenase from Landschütz ascites cells, calf thymus and LS cells are strongly inhibited by mycophenolic acid. The inhibition showed mixed type kinetics with K(i) values of between 3.03x10(-8) and 4.5x10(-8)m. 9. Evidence was also obtained for a partial, possibly indirect, inhibition by mycophenolic acid of an early stage of biosynthesis of purine nucleotides as indicated by a decrease in the accumulation of formylglycine amide ribonucleotide induced by the antibiotic azaserine in suspensions of Landschütz and Yoshida ascites cells and L cells in vitro.     

A COMPARISON OF THE CHANGES IN FINE STRUCTURE OF L CELLS DURING SINGLE CYCLES OF VIRAL MULTIPLICATION, FOLLOWING THEIR INFECTION WITH THE VIRUSES OF MENGO AND ENCEPHALOMYOCARDITIS

The virus of encephalomyocarditis (EMC), examined by the negative-contrast method, is indistinguishable from the serologically related Mengovirus. The particles are 270 to 280 A in diameter. The surface of EMC is composed of an undetermined number of subunits. Frequent sampling of infected cells was carried out throughout one-step cycles of viral multiplication to observe cytopathic changes occurring in L cells infected by these two related RNA viruses. EMC and Mengovirus, which multiply at equal rates, in most respects elicit similar alterations in cell fine structure. Rearrangement and changes in nuclear material accompanied by formation of small vesicles in the centrosphere region commence at 4 to 6 hours after infection. Thereafter a progressive degeneration of the nucleus and vesiculation of the cytoplasm are observed up to 18 to 20 hours. Increased numbers of small dense granules, indistinguishable from ribonucleoprotein particles, appear in the cytoplasm between 8 and 14 hours after infection. L cells infected with Mengovirus become permeable to Erythrocin more slowly than those infected with EMC. Only in the case of Mengovirus infection are large aggregates of dense material first observed in the cytoplasm at 8 hours, followed by the appearance of crystals probably composed of Mengovirus particles, at 12 hours. Differences in the rates of cell permeability after infection with EMC and Mengovirus are discussed in relation to formation of virus crystals and plaque-type mutants.