title: Induction of a new ribonucleotide reductase after infection of mouse L cells with pseudorabies virus.
abstract: The mammalian ribonucleotide reductase consists of two nonidentical subunits, protein M1 and M2. M1 binds nucleoside triphosphate allosteric effectors, whereas M2 contains a tyrosine free radical essential for activity. The activity of ribonucleotide reductase increased 10-fold in extracts of mouse L cells 6 h after infection with pseudorabies virus. The new activity was not influenced by antibodies against subunit M1 of calf thymus ribonucleotide reductase, whereas the reductase activity in uninfected cells was completely neutralized. Furthermore, packed infected cells (but not mock-infected cells) showed an electron paramagnetic resonance spectrum of the tyrosine free radical of subunit M2 of the cellular ribonucleotide reductase. These data given conclusive evidence that on infection, herpesvirus induces a new or modified ribonucleotide reductase. The virus-induced enzyme showed the same sensitivity to inhibition by hydroxyurea as the cellular reductase. The allosteric regulation of the virus enzyme was completely different from the regulation of the cellular reductase. Thus, CDP reduction catalyzed by the virus enzyme showed no requirement for ATP as a positive effector, and no feedback inhibition was observed by dTTP or dATP. The virus reductase did not even bind to a dATP-Sepharose column which bound the cellular enzyme with high affinity.
mesh_terms: D000494:Allosteric Regulation; D000818:Animals; D002846:Chromatography, Affinity; D004578:Electron Spin Resonance Spectroscopy; D011558:Herpesvirus 1, Suid / Q000201:enzymology*; D006918:Hydroxyurea / Q000494:pharmacology; D007739:L Cells / Q000032:analysis; D051379:Mice; D009500:Neutralization Tests; D012264:Ribonucleotide Reductases / Q000302:isolation & purification* / Q000378:metabolism; D013379:Substrate Specificity; D014764:Viral Proteins / Q000302:isolation & purification* / Q000378:metabolism
chemical_list: D014764:Viral Proteins; D012264:Ribonucleotide Reductases; D006918:Hydroxyurea